Methods and compositions for the sustained release of chromium

ABSTRACT

Provided herein are compositions for the administration of chromium that include at least two components: a hydrophilic chromium complex and a lipophilic chromium complex, and methods of using the same. Also provided are compositions for the administration of chromium that include a first “fast-acting” chromium complex and a second “slow-acting” chromium complex, wherein the first chromium complex is absorbed more quickly than the slow-acting chromium complex, and methods of using the same. Also provided herein are methods for treating, preventing, and improving conditions associated with cardiometabolic syndrome, by identifying a subject in need of treatment, prevention, or improvement of a condition associated with cardiometabolic syndrome, and providing a therapeutically effective amount of a composition comprising a fast-acting chromium complex and a slow-acting chromium complex, to the individual.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/620,542 filed on Jun. 12, 2017, now granted as U.S. Pat. No.10,245,325, which is a continuation of U.S. application Ser. No.14/805,136 filed on Jul. 21, 2015, now granted as U.S. Pat. No.9,675,702, which is a continuation of U.S. application Ser. No.13/300,059 filed on Nov. 18, 2011, now granted as U.S. Pat. No.9,119,835, which is a divisional of U.S. application Ser. No. 12/340,257filed on Dec. 19, 2008, now granted as U.S. Pat. No. 8,062,677, which isa continuation of International Application No. PCT/US2008/056545 filedon Mar. 11, 2008, which claims the benefit of U.S. ProvisionalApplication 60/894,601 filed on Mar. 13, 2007. The entire contents ofthese applications are incorporated herein by reference in theirentirety.

BACKGROUND OF THE INVENTION Field of the Invention

Embodiments disclosed herein relate to compositions that comprise,consist essentially of, or consist of a hydrophilic chromium complex anda lipophilic chromium complex, and uses thereof, as well as compositionsthat comprise, consist essentially of, or consist of a first and asecond chromium complex, wherein the first chromium complex is absorbedmore quickly than the second chromium complex, and uses thereof. Otherembodiments disclosed herein relate to the use of compositionscomprising, consisting essentially of, or consisting of chromium andhistidine, chromium histidinate complex, chromium trihistidinate,chromium polyhistidinate complex, chromium acetate, chromium chloride,or chromium nicotinate or combinations thereof, includingpharmaceutically acceptable salts, hydrates, solvates, or mixturesthereof in combination with another chromium complex for sustainedrelease of chromium. These compositions are useful in ameliorating avariety of conditions including cardiometabolic syndrome and relatedconditions, diseases, and disorders, improving glucose tolerance andglucose metabolism, treatment of insulin resistance, reducingpre-prandial and post-prandial glucose levels, reducing hyperglycemiaand stabilizing serum glucose, reducing free fatty acids, reducingcortisol levels, improvement in lipid profiles, and promoting weightloss.

Description of the Related Art Cardiometabolic Syndrome

Cardiometabolic syndrome (CMS) describes a constellation of maladaptivecardiovascular, renal, metabolic, prothrombotic, and inflammatoryabnormalities. CMS is recognized as a disease entity by the AmericanSociety of Endocrinology, National Cholesterol Education Program, andWorld Health Organization, and is characterized by various salientfeatures such as obesity, hypertension, dyslipidemia, impaired glucosetolerance, increase in inflammatory markers such as C-reactive protein(CRP), cytokines, tumor necrosis factor alpha (TNFα), interleukins 6 and10 (IL-6 and IL-10), changes in cell adhesion molecules, prothromboticand fibrinolytic changes, increase in oxidative stress and endothelialdysfunction. Juturu, 2006 DPG Medical Nutrition Therapy. Several of theconditions associated with CMS, e.g., obesity, hyperlipidemia, anddiabetes, play a causal role in atherosclerotic cardiovascular diseases,which currently account for a considerable proportion of mortality andmorbidity in developed, developing and underdeveloped societies.

Atherosclerosis

Atherosclerosis is a slowly progressive disease characterized by theaccumulation of cholesterol within the arterial wall. Without wishing tobe bound by any particular theory and solely for the purposes ofexpanding knowledge in the field, it is thought that lipids deposited inatherosclerotic lesions are derived primarily from plasma apolipoproteinB (apo B)-containing lipoproteins, which include chylomicrons, very lowdensity lipoproteins (VLDL), intermediate-density lipoproteins (IDL),and LDL. Apo B-containing lipoproteins, and in particular LDL, areassociated with adverse health outcomes. By contrast, HDL serum levelscorrelate inversely with coronary heart disease. Indeed, high serumlevels of HDL are regarded as a negative risk factor for CHD, andstudies suggest that high levels of plasma HDL are not only protectiveagainst coronary artery disease, but may actually induce regression ofatherosclerotic plaque. See, e.g., Badimon et al., 1992 Circulation86:(Suppl. III) 86 94; Dansky and Fisher, 1999, Circulation 100:1762 3.Data also suggest that non-HDL cholesterol (non HDL-C) might be a betterpredictive risk factor of CVD than LDL-C. The Adult Treatment Panel(ATP-Ill) recommended using non-HDL-C in assessing CVD risk in patientswith Type II Diabetes Mellitus.

Cholesterol

As discussed above, elevated serum cholesterol is linked to coronaryheart disease. Circulating cholesterol is carried by plasmalipoproteins, which are particles of complex lipid and proteincomposition that transport lipids in the blood. Low density lipoprotein(LDL) and high density lipoprotein (HDL) are the majorcholesterol-carrier proteins. LDL is believed to be responsible for thedelivery of cholesterol from the liver, where it is synthesized orobtained from dietary sources, to extrahepatic tissues in the body.“Reverse cholesterol transport” refers to the transport of cholesterolfrom extrahepatic tissues to the liver, where it is catabolized andeliminated. It is believed that plasma HDL particles play a major rolein the reverse transport process, acting as scavengers of tissuecholesterol. HDL is also responsible for the removal of non-cholesterollipid, oxidized cholesterol and other oxidized products from thebloodstream. The atherogenic index of plasma (AIP), defined as logarithm[log] of the ratio of plasma concentration of triglycerides (TG) toHDL-cholesterol (TG/HDL-C), has recently been proposed as a predictivemarker for plasma atherogenicity and is positively correlated withcardiovascular disease (CVD). Lipoprotein subclass abnormalities thataccompany insulin resistance are characterized by large,triglyceride-enriched very low-density lipoprotein (VLDL) particles;small, cholesterol-depleted LDL particles; and small HDL particles. Inaddition, more severe states of insulin resistance have been associatedwith progressively higher numbers of VLDL particles,intermediate-density lipoprotein particles and, most importantly, LDLparticles. The strong correlation of atherogenic index in plasma withlipoprotein particle size may explain its association withcardiovascular disease (CVD) risk. Atherogenic dyslipidemia results inincreased atherosclerotic plaque formation because of an imbalancebetween an increased number of small, dense LDL particles, which carrycholesterol to the vascular endothelium, and a decreased number of HDLparticles, which remove cholesterol from atherosclerotic vessels.Insulin resistance is the initial physiological defect in thepathogenesis of diabetes, such as Type II diabetes mellitus (“T2DM”);the associated atherogenic lipoprotein phenotype considerably enhancesthe risk of CVD. The combination of these factors may lead tocardiometabolic syndrome which is different from metabolic syndrome.Hyperinsulinemia is often clustered with other cardiovascular riskfactors; the presence of endogenous hyperinsulinemia combined withhypertriglyceridemia (HTG), increased body mass index, and a decreasedHDL-C increase the risk of CHD death in patients with T2DM. Castro etal, 2003, Curr Hypertens Rep. 5(5):393-401; Lastra et al. 2006, CurrDiab Rep. 6(3):207-12.

Cholesterol Transport

The fat-transport system can be divided into two pathways: an exogenousone for cholesterol and triglycerides absorbed from the intestine and anendogenous one for cholesterol and triglycerides entering thebloodstream from the liver and other non-hepatic tissue.

In the exogenous pathway, dietary fats are packaged into lipoproteinparticles called chylomicrons, which enter the bloodstream and delivertheir triglycerides to adipose tissue for storage and to muscle foroxidation to supply energy. The remnant of the chylomicron, whichcontains cholesteryl esters, is removed from the circulation by aspecific receptor found only on liver cells. This cholesterol thenbecomes available again for cellular metabolism or for recycling toextrahepatic tissues as plasma lipoproteins.

In the endogenous pathway, the liver secretes a large, very-low-densitylipoprotein particle (VLDL) into the bloodstream. The core of VLDLconsists mostly of triglycerides synthesized in the liver, with asmaller amount of cholesteryl esters either synthesized in the liver orrecycled from chylomicrons. Two predominant proteins are displayed onthe surface of VLDL, apolipoprotein B-100 (apo B-100) and apolipoproteinE (apo E), although other apolipoproteins are present, such asapolipoprotein CIII (apo CIII) and apolipoprotein CII (apo CII). WhenVLDL reaches the capillaries of adipose tissue or of muscle, itstriglyceride is extracted. This results in the formation of a new kindof particle called intermediate-density lipoprotein (IDL) or VLDLremnant, decreased in size and enriched in cholesteryl esters relativeto a VLDL, but retaining its two apoproteins.

In human beings, about half of the IDL particles are removed from thecirculation quickly, generally within two to six hours of theirformation. This is because IDL particles bind tightly to liver cells,which extract IDL cholesterol to make new VLDL and bile acids. The IDLnot taken up by the liver is catabolized by the hepatic lipase, anenzyme bound to the proteoglycan on liver cells. Apo E dissociates fromIDL as it is transformed to LDL. Apo B-100 is the sole protein of LDL.

Primarily, the liver takes up and degrades circulating cholesterol tobile acids, which are the end products of cholesterol metabolism. Theuptake of cholesterol-containing particles is mediated by LDL receptors,which are present in high concentrations on hepatocytes. The LDLreceptor binds both apo E and apo B-100 and is responsible for bindingand removing both IDL and LDL from the circulation. In addition, remnantreceptors are responsible for clearing chylomicrons and VLDL remnants,i.e., IDL. However, the affinity of apo E for the LDL receptor isgreater than that of apo B-100. As a result, the LDL particles have amuch longer circulating life span than IDL particles; LDL circulates foran average of two and a half days before binding to the LDL receptors inthe liver and other tissues. High serum levels of LDL are positivelyassociated with coronary heart disease. For example, in atherosclerosis,cholesterol derived from circulating LDL accumulates in the walls ofarteries. This accumulation forms bulky plaques that inhibit the flow ofblood until a clot eventually forms, obstructing an artery which mayultimately lead to heart attack or stroke.

Ultimately, the amount of intracellular cholesterol liberated from theLDL controls cellular cholesterol metabolism. The accumulation ofcellular cholesterol derived from VLDL and LDL controls three processes.First, it reduces the ability of the cell to make its own cholesterol byturning off the synthesis of HMGCoA reductase, a key enzyme in thecholesterol biosynthetic pathway. Second, the incoming LDL-derivedcholesterol promotes storage of cholesterol by the action of cholesterolacyltransferase (“ACAT”), the cellular enzyme that converts cholesterolinto cholesteryl esters that are deposited in storage droplets. Third,the accumulation of cholesterol within the cell drives a feedbackmechanism that inhibits cellular synthesis of new LDL receptors. Cells,therefore, adjust their complement of LDL receptors so that enoughcholesterol is brought in to meet their metabolic needs, withoutoverloading.

High levels of apo B-containing lipoproteins can be trapped in thesubendothelial space of an artery and undergo oxidation. The oxidizedlipoprotein is recognized by scavenger receptors on macrophages. Bindingof oxidized lipoprotein to the scavenger receptors can enrich themacrophages with cholesterol and cholesteryl esters independently of theLDL receptor. Macrophages can also produce cholesteryl esters by theaction of ACAT. LDL can also be complexed to a high molecular weightglycoprotein called apolipoprotein (a), also known as apo(a), through adisulfide bridge. The LDL-apo(a) complex is known as Lipoprotein(a) orLp(a). Elevated levels of Lp(a) are detrimental, having been associatedwith atherosclerosis, coronary heart disease, myocardial infarction,stroke, cerebral infarction, and restenosis following angioplasty. Wanget al. 2006, J Lipid Res. 5.

Reverse Cholesterol Transport

Peripheral (non-hepatic) cells predominantly obtain their cholesterolfrom a combination of local synthesis and uptake of preformed sterolfrom VLDL and LDL. Cells expressing scavenger receptors, such asmacrophages and smooth muscle cells, can also obtain cholesterol fromoxidized apo B-containing lipoproteins. In contrast, reverse cholesteroltransport (RCT) is the pathway by which peripheral cell cholesterol canbe returned to the liver for recycling to extrahepatic tissues, hepaticstorage, or excretion into the intestine in bile. The RCT pathwayrepresents the only means of eliminating cholesterol from mostextrahepatic tissues and is crucial to the maintenance of the structureand function of most cells in the body.

The enzyme in blood involved in the RCT pathway, lecithin:cholesterolacyltransferase (LCAT), converts cell-derived cholesterol to cholesterylesters, which are sequestered in HDL destined for removal. LCAT isproduced mainly in the liver and circulates in plasma associated withthe HDL fraction. Cholesterol ester transfer protein (CETP) and anotherlipid transfer protein, phospholipid transfer protein (PLTP), contributeto further remodeling the circulating HDL population. PLTP supplieslecithin to HDL, and CETP can move cholesteryl esters made by LCAT toother lipoproteins, particularly apoB-containing lipoproteins, such asVLDL. HDL triglycerides can be catabolized by the extracellular hepatictriglyceride lipase and lipoprotein cholesterol is removed by the livervia several mechanisms.

Each HDL particle contains at least one molecule, and usually two tofour molecules, of apolipoprotein A I (apo A I). Apo A I is synthesizedby the liver and small intestine as preproapolipoprotein, which issecreted as a proprotein that is rapidly cleaved to generate a maturepolypeptide having 243 amino acid residues. Apo A I consists mainly of a22 amino acid repeating segment, spaced with helix-breaking prolineresidues. Apo A I forms three types of stable structures with lipids:small, lipid-poor complexes referred to as pre-beta-1 HDL; flatteneddiscoidal particles, referred to as pre-beta-2 HDL, which contain onlypolar lipids (e.g., phospholipid and cholesterol); and sphericalparticles containing both polar and nonpolar lipids, referred to asspherical or mature HDL (HDL3 and HDL2). Most HDL in the circulatingpopulation contains both apo A I and apo A II, a second major HDLprotein. The apo A I- and apo A II-containing fraction is referred toherein as the AI/AII-HDL fraction of HDL. The fraction of HDL containingonly apo A I, referred to herein as the AI HDL fraction, appears to bemore effective in RCT. Certain epidemiologic studies support thehypothesis that the Al-HDL fraction is antiartherogenic. Spady et al.1999, Circulation. 100:576-578; Fielding C J, Fielding P E. 1995, JLipid Res. 36:211-228.

The LCAT reaction requires an apolipoprotein such as apo A I or apo A-IVas an activator. ApoA-I is one of the natural cofactors for LCAT. Theconversion of cholesterol to its HDL-sequestered ester prevents re-entryof cholesterol into the cell, resulting in the ultimate removal ofcellular cholesterol.

HDL is not only involved in the reverse transport of cholesterol, butalso plays a role in the reverse transport of other lipids, e.g., thetransport of lipids from cells, organs, and tissues to the liver forcatabolism and excretion. Such lipids include sphingomyelin, oxidizedlipids, and lysophophatidylcholine. For example, Robins and Fasulo haveshown that HDL stimulates the transport of plant sterol by the liverinto bile secretions. Robins and Fasulo (1997, J. Clin. Invest. 99:380384.

Cardiometabolic Syndrome

Available data suggest that insulin resistance is responsible for morethan 40% of heart attacks and is the underlying cause for various riskfactors, which lead to cardiometabolic syndrome (CMS).

In cardiovascular tissues there are two pathways of insulin receptorsignaling: one that is predominant in metabolic tissues (mediated byphosphatidylinositol-3-kinase); and a growth factor-like pathway(mediated by MAPK). The first pathway can lead to atherosclerosis.Patients presenting with multiple cardiometabolic risk factors havetriple the risk of experiencing a myocardial infarction and/or strokeand double the risk of mortality. In addition, the risk for developingtype 2 diabetes, if not already present, is fivefold above the risk inpatients without CMS.

In addition to the risks discussed above, hyperinsulinemia andhypertension, two conditions associated with CMS, can also contributesignificantly to progressive renal disease. Other mechanisms thatpotentially lead to progressive renal disease and CMS can includeendothelial dysfunction, left ventricular hypertrophy (LVH), cardiachyperreactivity, dyslipidemia, hyperglycemia, enhancedrenin-angiotensin-aldosterone system (RAAS) activity, altered renalstructure and function with impaired pressure natriuresis leading tosodium retention, volume expansion, progressive renal disease, andeventually end-stage renal disease (ESRD).

It has also been suggested that the impact of the CMS is largelyindependent from glycaemic control, and is associated with severalneglected modifiable and non modifable risk factors, such as abdominalobesity, especially visceral obesity. A common pathophysiologic process,such as endothelial dysfunction, chronic low-grade inflammation, orincreased transvascular leakage of macromolecules, can underlie theassociation between microalbuminuria and cardiovascular disease.Microalbuminuria has been implicated as an independent risk factor forCVD and premature cardiovascular mortality for patients with type 1 andtype 2 diabetes mellitus, as well as for patients with essentialhypertension. The combination of diabetes and CHD risk factors could beexplained by metabolic abnormalities that are not currently assessed indaily clinical practice. It is therefore suggested that in order tooptimally manage these risk factors, attention should be given not onlyto reduce risk factors, but also to the improvement of features of theCMS Juturu, 2006 DPGMNT.

Insulin resistance is a condition that is characterized by decreasedinsulin function and hyperinsulinemia. Individuals who have insulinresistance also have an increased risk of developing diabetes mellitus,dyslipidemia, hypertension, atherosclerosis, endothelial dysfunction,microalbuminuria, obesity, depression, Syndrome X, and polycystic ovarysyndrome, among other conditions. In addition, all of the aforementionedconditions carry the risk of developing associated diseases. Forexample, diabetes increases the risk of developing associated diseasessuch as diabetic nephropathy, neuropathy, and retinopathy.

Insulin resistance may result from taking certain drug therapies such asstatins, non-steroidal anti-inflammatory drugs (NSAIDS), steroids, oralcontraceptives, hormone replacement therapy (HRT), beta blockers,potassium channel openers, diuretics, immunosuppressive drugs, etc. Forexample, A. Jula et al. report that fasting serum insulin levelsincreased 13% and insulin resistance increased by 14% in 120 nondiabetichypercholesterolemic male patients taking statin drugs to reduce theircholesterol levels. A. Jula et al., 2002, JAMA 287:598-605, 604.Furthermore, it has also been reported that beta blockers and diureticsworsen insulin resistance and that patients taking beta blockers had a28% higher incidence of diabetes than untreated patients withhypertension. S. Julius et al., 2001, Am. J. Hypertens. 14:310S-316S,313S.

Insulin resistance has also been described as a side effect of a varietyof oral contraceptives. In a study of the metabolic effects ofimplantable steroid contraceptives, altered glucose tolerancecharacterized by decreased insulin sensitivity following glucoseadministration was seen in individuals with implantable contraceptives,such as NORPLANT®, JADELLE®, and IMPLANON® was observed. Dorfgliner, L.J., 2002, Contraception 65:47-62, Peterson, K. R., 2002, Danish MedicalBulletin 49:43-60. Similarly, oral contraceptives and hormonereplacement therapy (“HRT”) have been linked to the onset ofmicroalbuminuria. Monster, T. B. M et al., 2001, Arch Intern Med.161:2000-2005.

Often, physicians will prescribe a hypoglycemic drug such as metformin,which the patient must continue to take for the rest of the patient'slife, for individuals presenting with insulin resistance.

The Role of Chromium

Dietary supplementation of chromium to normal individuals has beenreported to lead to improvements in glucose tolerance, serum lipidconcentrations, including high-density lipoprotein cholesterol, insulinand insulin binding. Anderson, 1986 Clin. Psychol. Biochem. 4:31-41.Supplemental chromium in the trivalent form, e.g. chromic chloride, isassociated with improvements of risk factors associated with adult-onset(Type 2) diabetes and cardiovascular disease.

Chromium is a nutritionally essential trace element. The necessity ofdietary chromium was established in 1959 by Schwartz. Schwartz, “PresentKnowledge in Nutrition,” page 571, fifth edition (1984, the NutritionFoundation, Washington, D.C.). Chromium depletion is characterized bythe disturbance of glucose, lipid and protein metabolism and by ashortened lifespan. Chromium is essential for optimal insulin activityin all known insulin-dependent systems. Boyle et al., 1977 Southern Med.J. 70:1449-1453. Insufficient dietary chromium has been linked to bothmaturity-onset diabetes and to cardiovascular disease.

The principal energy sources for the body are glucose and fatty acids.Chromium depletion results in biologically ineffective insulin andcompromised glucose metabolism. Under these conditions, the body reliesprimarily upon lipid metabolism to meet its energy requirements, whichcan lead to the production of elevated amounts of acetyl-CoA and ketonebodies. In some cases, some of the acetyl-CoA can be diverted toincreased cholesterol biosynthesis, resulting in hypercholesterolemia.As such, glycosuria, hypercholesterolemia, and often ketoacidosis areoften associated with diabetes mellitus. The accelerated atheroscleroticprocess seen in diabetics is associated with hypercholesterolemia Boyleet al., supra.

Chromium functions as a cofactor for insulin. It binds to the insulinreceptor and potentiates many, and perhaps all, of its functions. Boyleet al., supra. These functions include, but are not limited to, theregulation of carbohydrate and lipid metabolism. Present Knowledge inNutrition, supra, at p. 573-577. The introduction of inorganic chromiumcompounds per se into individuals is not particularly beneficial.Chromium must be converted endogenously into an organic complex or mustbe consumed as a biologically active molecule. Only about 0.5% ofingested inorganic chromium, however, is assimilated into the body. Only1-2% of most organic chromium compounds are assimilated into the body.Recommended Daily Allowances, Ninth Revised Edition, The NationalAcademy of Sciences, page 160, 1980.

U.S. Pat. No. Re. 33,988 discloses that when selected essential metals,including chromium, are administered to mammals as exogenouslysynthesized coordination complexes of picolinic acid, they are directlyavailable for absorption without competition from other metals. U.S.Pat. No. Re. 33,988 describes a composition and method for selectivelysupplementing the essential metals in the human diet and forfacilitating absorption of these metals by intestinal cells. Thesecomplexes are safe, inexpensive, biocompatible, and easy to produce.These exogenously synthesized essential metal coordination complexes ofpicolinic acid (pyridine-2-carboxylic acid) have the followingstructural formula:

wherein M represents the metallic cation and n is equal to the cation'svalence. For example, when M is Cr and n=3, then the compound is chromictripicolinate. Other chromium picolinates disclosed include chromicmonopicolinate and chromic dipicolinate.

The U.S. Recommended Daily Intake (RDI) of chromium is 120 μg. U.S. Pat.No. 5,087,623, the entire contents of which are hereby expresslyincorporated herein by reference, describes the administration ofchromic tripicolinate for the treatment of adult-onset diabetes in dosesranging from 50 to 500 μg. U.S. Pat. No. 6,329,361, the entire contentsof which are hereby expressly incorporated herein by reference,discloses the use of high doses of chromic tripicolinate (providing1,000-10,000 μg chromium/day) for reducing hyperglycemia and stabilizingthe level of serum glucose in humans with Type 2 diabetes. U.S. Pat.Nos. 5,789,401 and 5,929,066, the entire contents of which are herebyexpressly incorporated herein by reference, disclose a chromictripicolinate-biotin composition and its use in lowering blood glucoselevels in humans with Type 2 diabetes.

U.S. Pat. Nos. 5,087,623; 5,087,624; and 5,175,156, the entire contentsof which are hereby expressly incorporated herein by reference, disclosethe use of chromium tripicolinate for supplementing dietary chromium,reducing hyperglycemia and stabilizing serum glucose, increasing leanbody mass and reducing body fat, and controlling serum lipid levels,including the lowering of undesirably high serum LDL-cholesterol levelsand the raising of serum High Density Lipid (HDL)-cholesterol levels.U.S. patent application Ser. Nos. 10/090,038 and 11/136,794, the entirecontents of which are hereby expressly incorporated by reference intheir entireties, disclose the use of high doses of chromium complexes(providing between 1,000 and 10,000 μg/day) and biotin for treatingdyslipidemia, and increasing serum HDL levels.

U.S. Pat. Nos. 4,954,492 and 5,194,615, the entire contents of which arehereby expressly incorporated by reference, describe a related complex,chromic nicotinate, which is also used for supplementing dietarychromium and lowering serum lipid levels. Picolinic acid and nicotinicacid are position isomers having the following structures:

Nicotinic acid and picolinic acid form coordination complexes withmonovalent, divalent and trivalent metal ions and facilitate theabsorption of these metals by transporting them across intestinal cellsand into the bloodstream.

Other compounds such as non-steroidal anti-inflammatory drugs (NSAIDs)such as aspirin and indomethachin have also been shown to facilitatechromium absorption. For Example, Davis et al. demonstrated that orallyadministered CrCl₃ is facilitated by the non-steroidal anti-inflammatorydrugs (NSAIDs) aspirin and indomethacin. Davis et al., 1995, J.Nutrition Res. 15:202-210 (1995); Kamath et al., 1997, J. Nutrition127:478-482. These drugs inhibit the enzyme cyclooxygenase whichconverts arachidonic acid to various prostaglandins, resulting ininhibition of intestinal mucus formation and lowering of intestinal pHwhich facilitates chromium absorption.

U.S. Pat. No. 4,315,927 teaches that when selected essential metals areadministered to mammals as exogenously synthesized coordinationcomplexes of picolinic acid, they are directly available for absorptionwithout competition from other metals. These complexes are safe,inexpensive, biocompatible and easy to produce.

There remains a need for sources of chromium that exhibit favorableabsorption profiles, and also that provide for the release of chromiumfrom the coordination complex once within the cell. Further, thereremains a need for compositions that provide sources of chromium withdiffering absorption profiles to provide a fast and slow-acting sourceof chromium.

SUMMARY OF THE INVENTION

Provided herein are improved compositions for supplementing chromium inindividuals. Some embodiments provide compositions for theadministration of chromium over a period of time. Such compositions cancomprise, consist of, or consist essentially of a lipophilic chromiumcomplex and a hydrophilic chromium complex, or a first chromium complexand a second chromium complex, wherein the first chromium complex isabsorbed more quickly (fast-acting) than the second chromium complex(slow-acting). For example, in some embodiments, the lipophilic chromiumcomplex or slow-acting chromium complex can be chromium picolinate orchromium tripicolinate, and the hydrophilic chromium complex orfast-acting chromium complex can be any one of chromium acetate,chromium chloride, chromium histidinate, and chromium nicotinate, or anycombination thereof. In some embodiments, the hydrophilic chromiumcomplex or fast-acting chromium complex is chromium histidinate. In someembodiments, the slow-acting or lipophilic chromium complex is chromiumpicolinate. In some embodiments, the compositions are pharmaceuticalcompositions comprising one or more compositions disclosed herein, witha pharmaceutically acceptable vehicle, excipient, or diluent. Forexample, pharmaceutically acceptable vehicles can include carriers,excipients, diluents, and the like, as well as combinations or mixturesthereof.

Also provided herein are methods of using the compositions describedherein. For example, embodiments provide methods for reducingpre-prandial and post-prandial glucose levels, for reducinghyperglycemia or stabilizing serum glucose levels, for improving insulinsensitivity, for reducing free fatty acid levels, and for treatingdyslipidemia by identifying a subject in need of a reduction in pre orpost-prandial glucose levels, a reduction in hyperglycemia, astabilization of serum glucose levels, and improvement in insulinsensitivity, a reduction in free fatty acid levels, or treatment fordyslipidemia, and providing a therapeutically effective amount of thecompositions described above to said individual. In some embodiments,the therapeutically effective amount of the fast-acting and theslow-acting chromium complex provides a glucose lowering effect orglucose stabilization effect for at least 10 hours.

In certain embodiments of the methods described herein, the hydrophilicchromium complex and the lipophilic chromium complex, or the firstchromium complex and the second chromium complex can be provided atsubstantially the same time, e.g., in a single composition. In otherembodiments, the hydrophilic chromium complex and the lipophilicchromium complex, or the first chromium complex and the second chromiumcomplex can be provided sequentially in either order.

Embodiments disclosed herein also relate to improved compositions forlowering serum glucose levels, for improving insulin sensitivity, fortreating dyslipidemia, and for increasing lean muscle mass. The improvedcompositions can include synergistically effective amount of ahydrophilic and a lipophilic chromium complex, e.g., chromiumhistidinate and chromium picolinate, wherein the synergisticallyeffective amount of chromium histidinate and said chromium picolinatehave a greater than additive effect in lowering serum glucose levels,improving insulin sensitivity, treating dyslipidemia, and increasinglean muscle mass, respectively.

In some embodiments, the ratio of the lipophilic or slow-acting chromiumcomplex to the hydrophilic or fast-acting chromium complex is about0.0001:1, 0.001:1, 0.01:1, 0.1:1, 0.2:1; 0.3:1, 0.4:1; 0.5:1; 0.75:1,1:1, 1:1.1, 1.2:1, 1.3:1, 1.4:1, 1.5:1, 1.75:1, 2:1, 5:1, 10:1, 100:1,500:1, or any ratio in between.

Also provided herein are compositions comprising chromium and histidine,chromium histidinate, chromium histidinate complexes, and combinationsthereof, e.g., chromium with histidinate or histidinate complex or polyhistidinate or mono histidinate in combination with a lipophilicchromium complex, such as chromium picolinate. In certain embodiments,the compositions described herein can be used in combination with othertherapeutics, such as hypocholesterolemic and hypoglycemic therapeuticagents.

Compositions including chromium and histidine, chromium histidinate,chromium histidinate complexes, and combinations thereof in combinationwith lipophilic or slow-acting chromium complexes disclosed hereinprovide unexpected benefits over different sources of chromium,including various known chromium complexes, in the treatment andprevention a variety of diseases and conditions such as, but not limitedto, aging, Alzheimer's Disease, cancer, cardiovascular disease, diabeticnephropathy, diabetic retinopathy, a disorder of glucose metabolism,disorders of lipid metabolism, dyslipidemia, dyslipoproteinemia,hypertension, impotence, inflammation, insulin resistance, lipidelimination in bile, obesity, oxysterol elimination in bile,pancreatitis, Parkinson's disease, a peroxisome proliferator activatedreceptor-associated disorder, renal disease, septicemia, Syndrome X,thrombotic disorder, and the like. Compositions and methods describedherein can also be used to modulate C-reactive protein or enhance bileproduction in a mammal, and eliminate or reduce phospholipid in bile.

Accordingly, provided herein are methods of treating or preventingcardiometabolic syndrome or a condition associated therewith in asubject that has or is at risk of developing CMS or a conditionassociated therewith, by providing the subject a composition thatcontains a fast-acting or hydrophilic chromium complex in combinationwith at least one slow-acting or lipophilic chromium complex. Forexample, in some embodiments, an individual that has or is at risk ofdeveloping CMS or a condition associated therewith is identified, andprovided a composition containing an effective dose of chromiumhistidinate and/or at least one other chromium complex, e.g., chromiumpicolinate.

Also provided herein are methods for inhibiting hepatic fatty acid andsterol synthesis in subjects in need thereof, by identifying subjects inneed of inhibition of hepatic fatty acids or inhibition of sterol, andproviding a therapeutically effective amount of the compositionsdisclosed herein to the subject. The administration or provision of afast-acting or hydrophilic chromium complex, e.g., chromium histidinateand a slow-acting or lipophilic chromium complex, e.g., chromiumpicolinate, for the prevention or inhibition of fatty acid and sterolsynthesis is also provided.

Also provided are methods for increasing HDL levels in a subject in needof increased HDL levels, by identifying a subject in need thereof, andproviding a therapeutically effective amount of a composition disclosedherein. Accordingly, embodiments disclosed herein also relate to thetreatment or prevention of diseases or disorders capable of beingtreated or prevented by increasing HDL levels in subjects in needthereof. For example a subject in need of increased HDL levels, e.g., asubject may believe it is in need or may self-identify or be identifiedusing routine methods, can be provided a composition comprising,consisting essentially of, or consisting of a fast-acting or hydrophilicchromium complex, e.g., chromium histidinate and a slow-acting orlipophilic chromium complex, e.g., chromium picolinate, for increasingHDL levels.

Provided herein are methods for lowering LDL levels in subjects in needof a reduction in LDL levels by identifying a subject in need of areduction in LDL levels, and providing a therapeutically effectiveamount of a composition disclosed herein to said subject. For example, asubject in need of decreased LDL levels can be provided a fast-acting orhydrophilic chromium complex, e.g., chromium histidinate, and aslow-acting or lipophilic chromium complex, e.g., chromium picolinate,for decreasing LDL levels.

Further provided herein are methods of improving endothelial function ina subject in need of improved endothelial function by identifying asubject in need of improved endothelial function, e.g., byself-identification or by routine clinical methods, and providing atherapeutically effective amount of a composition disclosed herein tosaid subject. For example, a subject in need of improved endothelialfunction can be provided a fast-acting or hydrophilic chromium complex,e.g., chromium histidinate, and a slow-acting or lipophilic chromiumcomplex, e.g., chromium picolinate, for improving endothelial function.

Disclosed herein are methods for improving at least one of thefollowing: blood pressure, vascular tone, vascular relaxation, andcoronary blood flow in a subject in need thereof by identifying asubject in need of improved blood pressure, vascular tone, vascularrelaxation, and coronary blood flow using routine clinical methods. Thesubject can be provided a therapeutically effective amount of acomposition disclosed herein. For example a subject in need of improvedblood pressure, vascular tone, vascular relaxation, and coronary bloodflow can be provided a fast-acting or hydrophilic chromium complex,e.g., chromium histidinate and a slow-acting or lipophilic chromiumcomplex, e.g., chromium picolinate, for improving blood pressure,vascular tone, vascular relaxation, and coronary blood flow.

Also provided are methods for lowering fasting and post prandial bloodsugar levels, lowering serum triglyceride levels and improving insulinsensitivity in a subject in need thereof by identifying a subject inneed of lower fasting and post-prandial blood sugar levels and providinga therapeutically effective amount of a composition disclosed herein tosaid subject. For example a subject in need of lower fasting andpost-prandial blood sugar levels can be provided a composition thatcomprises chromium and histidine, chromium histidinate complexes, orcombinations thereof in combination with at least one other slow-actingchromium complex. The administration or provision of a fast-acting orhydrophilic chromium complex, e.g., chromium histidinate and aslow-acting or lipophilic chromium complex, e.g., chromium picolinate,for lowering fasting and post-prandial blood sugar levels is alsoprovided.

The compositions disclosed herein can improve fasting and post prandialblood insulin levels, decrease hyperinsulinemia, and decrease insulinresistance in mammals. Accordingly, some embodiments provide methods fortreatment or prevention of cardiometabolic syndrome-associateddisorders, such as hyperglycemia, hyperinsulinemia, or insulinresistance. A subject in need of improved fasting and post-prandialblood insulin levels, a decrease in hyperinsulinemia, or a decrease ininsulin resistance may believe it is in need or may self-identify or beidentified using routine methods, and is provided a therapeuticallyeffective amount of a fast-acting and slow-acting chromium complex asdisclosed herein. For example, a subject in need of improved fasting andpost-prandial blood insulin levels, a decrease in hyperinsulinemia, or adecrease in insulin resistance can be provided a composition thatcontains chromium and histidine, chromium histidinate complexes, orcombinations thereof in combination with at least one slow-acting orlipophilic chromium complex. The administration or provision of afast-acting or hydrophilic chromium complex, e.g., chromium histidinateand a slow-acting or lipophilic chromium complex, e.g., chromiumpicolinate, for improving fasting and post-prandial blood insulinlevels, decreasing hyperinsulinemia, and decreasing insulin resistanceis also provided.

The compositions disclosed herein can decrease body fat and increaselean body mass, thereby effectuating improvements in body composition inmammals. Accordingly, some embodiments provide methods for decreasingbody fat or increasing lean body mass in a subject by identifying asubject in need of a decrease in body fat or increase in lean body mass,and providing to said subject a therapeutically amount of a fast-actingchromium complex in combination with a slow-acting chromium complex. Forexample a subject in need of a decrease in body fat or an increase inlean muscle mass can be provided a composition that comprises chromiumand histidine, chromium histidinate complexes, or combinations thereofin combination with at least one slow-acting or lipophilic chromiumcomplex. The administration or provision of a fast-acting or hydrophilicchromium complex, e.g., chromium histidinate and a slow-acting orlipophilic chromium complex, e.g., chromium picolinate, for decreasingbody fat or increasing lean muscle mass is also provided.

The compositions disclosed herein can decrease inflammatory markers, therisk of CVD and diabetes, and reduce obesity in mammals. Accordingly,some embodiments provide methods of decreasing inflammatory markers,decreasing the risk of CVD and diabetes, or reducing obesity in mammalsby identifying a subject in need of a decrease in inflammatory markers,a subject at risk of CVD and diabetes, or a subject that is obese, usingroutine clinical methods, and providing the subject a therapeuticallyeffective amount of a fast-acting chromium complex in combination with aslow-acting chromium complex. The administration or provision of afast-acting or hydrophilic chromium complex, e.g., chromium histidinateand a slow-acting or lipophilic chromium complex, e.g., chromiumpicolinate, for decreasing inflammatory markers is also provided.

The compositions disclosed herein decrease markers associated with renalfunction and help in improving renal function in mammals. Accordingly,provided herein are methods for the treatment or prevention of renaldisorders, by identifying a subject with or at risk of developing arenal disorder, e.g., a subject with cardiometabolic syndrome and arenal disorder, and providing a therapeutically effective amount of acomposition that contains chromium and histidine, chromium histidinatecomplexes, or combinations thereof in combination with at least oneslow-acting or lipophilic chromium complex. The administration orprovision of a fast-acting or hydrophilic chromium complex, e.g.,chromium histidinate and a slow-acting or lipophilic chromium complex,e.g., chromium picolinate, for improving renal function is alsoprovided.

The compositions disclosed herein can decrease inflammatory markersassociated with bone health and help in improving bone health anddisorders, for example in mammals. Accordingly, some embodiments providemethods of treatment or prevention of cardiometabolic syndrome disorderswith arthritis and rheumatic heart disease by identifying a subject withCMS that has or is at risk of developing arthritis or rheumatic heartusing routine clinical methods, and providing the subject atherapeutically effective amount of a fast-acting or hydrophilicchromium complex, e.g., chromium histidinate and a slow-acting orlipophilic chromium complex, e.g., chromium picolinate.

The compositions disclosed herein can improve immune function associatedwith cardiometabolic syndrome, for example in mammals. Accordingly,provided herein are methods for treating or preventing immune functiondisorders, by identifying a subject with CMS that is in need of improvedimmune function and providing the subject a therapeutically effectiveamount of a fast-acting or hydrophilic chromium complex, e.g., chromiumhistidinate and a slow-acting or lipophilic chromium complex, e.g.,chromium picolinate.

The compositions disclosed herein can improve metabolic functionassociated with cardiometabolic syndrome, diabetes, obesity andcardiovascular disease, for example in mammals. Accordingly, someembodiments provide methods of treatment or prevention ofcardiometabolic syndrome disorders with metabolic disorders. Anindividual with CMS, diabetes, obesity or cardiovascular disease andpoor metabolic function may believe it is in need or may self-identifyor be identified using routine methods, and provided a therapeuticallyeffective amount of a fast-acting or hydrophilic chromium complex, e.g.,chromium histidinate and a slow-acting or lipophilic chromium complex,e.g., chromium picolinate.

The compositions disclosed herein can improve chromium status associatedwith cardiometabolic syndrome, diabetes, obesity and cardiovasculardisease but not limited to other chronic conditions alone or incombination with other disorders, for example in mammals. Accordingly,some embodiments provide methods of treatment or prevention ofcardiometabolic syndrome disorders with low chromium status ordeficiency of chromium and methods of improving chromium depletion intissues due to chronic conditions, by identifying s subject with CMSwith low chromium status, and providing the subject a therapeuticallyeffective amount of a fast-acting or hydrophilic chromium complex, e.g.,chromium histidinate and a slow-acting or lipophilic chromium complex,e.g., chromium picolinate.

The compositions disclosed herein can improve amino acid profile statusand chromium absorption associated with cardiometabolic syndrome,diabetes, obesity and cardiovascular disease, for example in mammals.Accordingly, provided herein are methods of treatment or prevention ofcardiometabolic syndrome disorders in subjects with low amino acidprofile, deficiency of protein, or deficiency of all amino acids and toimprove amino acid profile depletion by identifying a subject with CMSand low amino acid profile, protein deficiency, and/or low chromiumstatus or levels, and providing the subject a therapeutically effectiveamount of a fast-acting or hydrophilic chromium complex, e.g., chromiumhistidinate and a slow-acting or lipophilic chromium complex, e.g.,chromium picolinate.

The compositions disclosed herein can improve exchange and transport ofchromium levels in the tissues associated with conditions such ascardiometabolic syndrome, diabetes, obesity and cardiovascular diseasein mammals and therefore the invention also encompasses methods ofimproving amino acid exchange and transport of amino acids for normalfunctions of the organs in the body by administering to the subject atherapeutically effective dose of a fast-acting or hydrophilic chromiumcomplex, e.g., chromium histidinate and a slow-acting or lipophilicchromium complex, e.g., chromium picolinate. Further provided aremethods for improving amino acid profile or deficiency of protein or allamino acids, methods for improving amino acid profile depletion, andmethods for improving amino acid absorption due to chronic conditionsand to replete the amino acids levels in tissues by administering to thesubject a therapeutically effective dose of a fast-acting or hydrophilicchromium complex, e.g., chromium histidinate and a slow-acting orlipophilic chromium complex, e.g., chromium picolinate.

The compositions disclosed herein improve exchange and transport ofchromium levels in the tissues associated with conditions such as aging,and chronic diseases such as cardiometabolic syndrome, diabetes, obesityand cardiovascular disease in mammals. Accordingly, some embodimentsrelate to methods improving chromium exchange along with other nutrientsand transport of chromium for normal functions of the organs in the bodyto the tissues and to improve chromium status or deficiency of chromiumor to improve chromium levels in tissues such as hair, skin, toenails,serum, blood, plasma and enhancing tissue concentrations in the organsincluding brain, heart, lung, liver, kidneys, spleen, and aorta.Preferably, the methods for improving chromium status and repletingchromium in the tissues and improving chromium levels or status inphysiology and biochemistry due to chronic conditions by administering atherapeutically effective dose of a fast-acting or hydrophilic chromiumcomplex, e.g., chromium histidinate and a slow-acting or lipophilicchromium complex, e.g., chromium picolinate.

The compositions disclosed herein favorably alter lipid metabolism inmammals with dyslipidemia at least in part by enhancing oxidation offatty acids through the ACC/malonyl-CoA/CPT-I regulatory axis.Accordingly, provided herein are methods of treating dyslipidemia byidentifying a subject with a poor lipid profile, and providing atherapeutically effective amount of a composition described herein tothe subject. For example, an individual with dyslipidemia can beprovided a a fast-acting or hydrophilic chromium complex, e.g., chromiumhistidinate and a slow-acting or lipophilic chromium complex, e.g.,chromium picolinate. The fast-acting or hydrophilic chromium complex canbe formulated with the slow-acting or lipophilic chromium complex.Alternatively, the fast-acting and slow-acting chromium complexes can beadministered separately.

Further embodiments provide methods for reducing the abdominal fat insubjects in need thereof by identifying subject in need of abdominalfat-content reduction and providing the subject a therapeuticallyeffective amount of a fast-acting or hydrophilic chromium complex, e.g.,chromium histidinate and a slow-acting or lipophilic chromium complex,e.g., chromium picolinate.

Provided herein are methods for reducing the total serum cholesterol ofa subject. A subject in need of a reduction in serum cholesterol levelsmay believe it is in need or may self-identify or be identified, andprovided a therapeutically effective amount of a a fast-acting orhydrophilic chromium complex, e.g., chromium histidinate and aslow-acting or lipophilic chromium complex, e.g., chromium picolinate,for lowering cholesterol levels is provided.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a bar graph showing the difference in glucose uptake byunstimulated and insulin-stimulated skeletal muscle cells cultured inthe presence of chromium picolinate or the combination of chromiumpicolinate/chromium histidinate (1:1) compared to untreated cultures.

FIGS. 2A and 2B are bar graphs showing the change in serum glucoselevels (mg/dL) in rats fed either a normal diet (16A) or a high fat diet(16B), supplemented with chromium picolinate (CrPic), chromiumhistidinate (CrHis), or a combination of chromium picolinate/chromiumhistidinate (CrPic:His) relative to rats that did not receive chromiumsupplementation.

FIGS. 3A and 3B are bar graphs showing the change in serum cholesterollevels (mg/dL) in rats fed either a normal diet (17A) or a high fat diet(17B), supplemented with chromium picolinate (CrPic), chromiumhistidinate (CrHis), or a combination of chromium picolinate/chromiumhistidinate (CrPic:His) relative to rats that did not receive chromiumsupplementation.

FIGS. 4A and 4B are bar graphs showing the change in serum triglyceridelevels (mg/dL) in rats fed either a normal diet (18A) or a high fat diet(18B), supplemented with chromium picolinate (CrPic), chromiumhistidinate (CrHis), or a combination of chromium picolinate/chromiumhistidinate (CrPic:His) relative to rats that did not receive chromiumsupplementation.

FIGS. 5A and 5B are bar graphs showing the change in free fatty acidlevels (mmol/L) in rats fed either a normal diet (18A) or a high fatdiet (18B), supplemented with chromium picolinate (CrPic), chromiumhistidinate (CrHis), or a combination of chromium picolinate/chromiumhistidinate (CrPic:His) relative to rats that did not receive chromiumsupplementation.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

The present invention is based, in part, on the novel and unexpecteddiscovery that the combination of a slow-acting, lipophilic chromiumcomplex with a fast-acting, hydrophilic chromium complex providesunexpected and improved properties when compared to either a fast-actingor a slow-acting chromium complex alone. Accordingly, described hereinare compositions that provide for improved administration of chromium toan individual, and methods of uses thereof, that include at least oneslow-acting, lipophilic chromium complex and at least one fast-acting,hydrophilic chromium complex.

In some embodiments, the compositions can comprise, consist essentiallyof, or consist of at least two components: a hydrophilic chromiumcomplex and a lipophilic chromium complex. In other embodiments, thecompositions can include a first chromium complex and a second chromiumcomplex, wherein the first chromium complex is absorbed more quickly(“fast-acting”) than the second chromium complex (“slow-acting”).

As used herein, the phrase “over a period of time,” can refer to aperiod of minutes, hours or days. For example, over a period of time canrefer to at least 10 minutes, at least 15 minutes, at least 30 minutes,at least 60 minutes, at least 75 minutes, at least 90 minutes, at least105 minutes, at least 120 minutes, at least 3 hours, at least 4 hours,at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours,at least 9 hours, at least 10 hours, at least 12 hours, at least 14hours, at least 16, hours, at least 18 hours, at least 20 hours, atleast 22 hours, at least one day, at least two days, at least threedays, at least 4 days, at least 5 days, at least 6 days, at least aweek, or any interval of time in between. In other words, the chromiumfrom the composition can be absorbed by the individual to whom it isadministered over a period of at least 10 minutes, at least 15 minutes,at least 30 minutes, at least 60 minutes, at least 75 minutes, at least90 minutes, at least 105 minutes, at least 120 minutes, at least 3hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7hours, at least 8 hours, at least 9 hours, at least 10 hours, at least12 hours, at least 14 hours, at least 16, hours, at least 18 hours, atleast 20 hours, at least 22 hours, at least one day, at least two days,at least three days, at least 4 days, at least 5 days, at least 6 days,at least a week, or any interval of time in between.

In some embodiments, the compositions provide bioavailable chromium fora period of time that is about the same as the period of time that a“slow-acting” chromium complex alone provides bioavailable chromium. Theterm “bioavailable,” in reference to chromium as used herein is meant torefer to the amount of chromium that is absorbed by an individual. Inpreferred embodiments, the compositions provide bioavailable chromiumover a total period of time that is greater than the period of time asthat of the “slow-acting” chromium complex alone provides bioavailablechromium. For example, the compositions described herein can providebioavailable chromium for a period of time that is at least 10 minutes,at least 15 minutes, at least 30 minutes, at least 60 minutes, at least75 minutes, at least 90 minutes, at least 105 minutes, at least 120minutes, at least 3 hours, at least 4 hours, at least 5 hours, at least6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least10 hours, at least 12 hours, at least 14 hours, at least 16, hours, atleast 18 hours, at least 20 hours, at least 22 hours, at least one day,at least two days, at least three days, at least 4 days, at least 5days, at least 6 days, at least a week, or any interval of time inbetween, greater than the “slow-acting” chromium complex alone. Byadministrating a slow-acting chromium complex together with afast-acting chromium complex, we have been able to achieve a sustainedrelease and absorption of chromium over a period of time.

As used herein, the term “hydrophilic chromium complex” or “fast actingchromium complex” refers to a chromium complex that is charged atphysiological pH, or has polar properties. Non-limiting examples ofhydrophilic, fast-acting chromium complexes include chromium acetate,chromium chloride, chromium histidinate and chromium nicotinate, and thelike, or any pharmaceutically acceptable salts, hydrates, solvates, ormixtures thereof.

The term “lipophilic chromium complex” or “slow-acting chromium complex”refers to a chromium complex that is not charged at physiological pH,and that does not have polar properties. Chromium picolinate, and anypharmaceutically acceptable salts, hydrates, or solvates thereof, is anon limiting example of a lipophilic, slow-acting chromium complex.

In preferred embodiments, the hydrophilic chromium complex or the“fast-acting” chromium complex is chromium histidinate, chromiumtrihistidinate, or chromium polyhistidinate, or any combination thereof.Preferably, the lipophilic chromium complex or the “slow-acting”chromium complex is chromium picolinate.

The terminology used in the description presented herein is not intendedto be interpreted in any limited or restrictive manner, simply becauseit is being utilized in conjunction with a detailed description ofcertain specific embodiments described herein. Furthermore, embodimentsdescribed herein can include several novel features, no single one ofwhich is solely responsible for its desirable attributes or which isessential to practicing the invention herein described. As used herein,the term “slow-acting chromium complexes” or “slow-acting chromiumcomplex” includes trivalent chromium complexes, such as chromiumpicolinate, chromic tripicolinate, and other lipophilic chromiumcomplexes, whether now known or developed in the future.

The present invention is based, in part, on the novel and unexpecteddiscovery that when a subject is administered a fast-acting chromiumcomplex and a slow-acting chromium complex, the symptoms and incidenceof insulin resistance are lowered. Accordingly, some embodiments providea method for the inhibition or reduction of insulin resistance, orlowering the risk of developing insulin resistance or related symptomsby lowering glucose and lipids and improving insulin sensitivity byadministering a fast-acting and a slow-acting chromium complex, e.g.,chromium histidinate and chromium picolinate.

As used herein, the term “altering lipid metabolism” indicates anobservable (measurable) change in at least one aspect of lipidmetabolism, including but not limited to total blood lipid content,blood HDL cholesterol, blood LDL cholesterol, blood VLDL cholesterol,blood triglyceride, blood Lp(a), blood apo A-I, blood apo E and bloodnon-esterified fatty acids, esters of fatty acids, isomers, isoforms andratios and improving ratios for reducing chronic disease risk but notlimited to diabetes, obesity, hypertension, coronary heart disease andcardiovascular disease.

As used herein, the term “altering glucose metabolism” indicates anobservable (measurable) change in at least one aspect of glucosemetabolism, including but not limited to total blood glucose content,blood insulin, the blood insulin to blood glucose ratio, glycosylatedhemoglobin, HOMAIR, beta cell function, composite of insulin sensitivityindex, hyperglycemia, hyperglycemia, hypoglycemia, hormones, enhancingenzyme activities, improving hormonal balance caused due to insulinresistance, abnormal glucose metabolism, lipodystrophy, reducing braininsulin resistance, insulin sensitivity, and oxygen consumption.Abnormal glucose metabolism in conditions like polycystic ovarysyndrome, HIV, HIV lipodystrophy, Alzheimer's disease, mental healthdisorders, lipodystrophy, hormonal imbalance conditions, hypertension,obesity and cardiovascular disease and cardiometabolic syndrome.

“Insulin resistance” refers to a condition characterized by decreasedinsulin function and hyperinsulinemia, caused or exacerbated by drugsand disease conditions such to obesity, diabetes, CVD in a human orother animal. Examples of drugs which induce insulin resistance include,without limitation, statin drugs such as simvastatin, cerivastatin,pravastatin, atorvastatin, fluvastatin, and lovastatin; non-steroidalanti-inflammatory drugs such as cimicifuga, choline salicylate-magnesiumsalicylate, diclofenac sodium, diclofenac potassium, diflunisal,etodolac, fenoprofen calcium, floctafenine, flurbiprofen, ibuprofen,indomethacin, ketoprofen, ketorolac tromethamine, magnesium salicylate,mefenamic acid, nabumetone, naproxen, naproxen sodium, oxyphenbutazone,phenylbutazone, piroxicam, salsalate, sodium salicylate, sulindac,tenoxicam, taiprofenic acid, and tolmetin sodium; steroids such ashydrocortisone, dexamethasone, and methylprednisolone; contraceptivesincluding oral contraceptives such as estrogen, progesterone andprogestin as well as implantable contraceptives such as levonorgestrel,etonogestrel, nomegestrol acetate, and nestorone; hormone replacementtherapy (HRT) drugs including conjugated equine estrogens, esterifiedestrogens, estradiol, estrone, synthetic conjugated estrogens,estropipate, estropipate, ethinyl estradiol, norethindrone,medroxyprogesterone acetate, progestin, natural progesterone, tamoxifen,testosterone, and raloxifene; beta blocker drugs including acebutolol,atenolol, betaxolol, bucinodol, carteolol, labetalol, metoprolol,nadolol, penbutolol, pindolol, propanolol, and timolol; and diuretics.Three primary types of diuretics exist which include thiazides, loopdiuretics, and potassium sparing agents. As used herein, the term“diuretic” or “diuretics” includes, without limitation,hydrochlorothiazide, chlorthalidone, chlorothiazide, indapamide,metolazone, amiloride, spironolactone, triamterene, furosemide,bumetanide, ethacrynic acid, and torsemide. Certain immunosuppressivedrugs such as prednisolone, cyclosporin A, and tacromlimus and potassiumchannel modulators such as nicorandil are also included in thedefinition of drugs which induce insulin resistance, such as, forexample, antidepressants. The above list is provided for examplepurposes only and it is understood that the definition of “drug whichinduces insulin resistance” includes those drugs which induce insulinresistance that are not specifically listed above, as well as thosedrugs which are found to induce insulin resistance, whether in existencetoday or developed in the future. Examples of diet which induce insulinresistance include diets high in fats, carbohydrates, low dietary fiber,high glycemic index foods, high fructose in the functional foods,beverages, and bars.

Other embodiments relate to the use of compositions comprising,consisting essentially of, or consisting of chromium and histidine,chromium histidinate complex, chromium trihistidinate, or chromium polyhistidinate complex, or combinations thereof, including pharmaceuticallyacceptable salts, hydrates, solvates, or mixtures thereof in combinationwith a second slow-acting chromium complex for the treatment orprevention of cardiometabolic syndrome and related conditions, diseases,and disorders.

The term “treating” or “treatment” does not necessarily mean total cure.Any alleviation of any undesired signs or symptoms of the disease to anyextent or the slowing down of the progress, or even prevention of thedisease or condition can be considered treatment.

Other embodiments relate to the use of compositions comprising,consisting essentially of, or consisting of chromium and histidine,chromium histidinate complex, chromium trihistidinate, or chromium polyhistidinate complex, or combinations thereof, including pharmaceuticallyacceptable salts, hydrates, solvates, or mixtures thereof in combinationwith a second slow-acting chromium complex for the maintenance ofhealthy or normal serum glucose levels, serum cholesterol levels, serumtriglyceride levels, or free fatty acid levels and/or the prevention ofthe development of hyperglycemia, dyslipidemia, high cholesterol, hightriglycerides, or high free fatty acid levels. In some embodiments, thesubject may have normal blood glucose, cholesterol, triglyceride, orfree fatty acid levels. Accordingly, some embodiments relate tomaintenance of fasting plasma glucose levels at less than about 100mg/dL (5.55 mmol per liter) in a subject, for example, in a healthysubject. Some embodiments relate to maintenance of total serumcholesterol levels below about 200 mg/dL, maintenance of serum HDLlevels above about 40-50 mg/dL; maintenance of serum LDL levels belowabout 100 mg/dL, maintenance of serum triglycerides at below about 150mg/dL, or the like.

The Role of Histidine/Histidinate

Histidine is one of the 20 most common natural amino acids present inproteins. In the nutritional sense, in humans, histidine is consideredan essential amino acid for normal healthy function. The imidazole sidechains and the relatively neutral pKa of histidine (ca 6.0) mean thatrelatively small shifts in cellular pH will change its charge. For thisreason, this amino acid side chain finds its way into considerable useas a coordinating ligand in metalloproteins, and also as a catalyticsite in certain enzymes. The imidazole side chain has two nitrogens withdifferent properties: one is bound to hydrogen and donates its lone pairto the aromatic ring and as such is slightly acidic; the other onedonates only one electron to the ring so it has a free lone pair and isbasic. These properties are exploited in different ways in proteins. Incatalytic triads, the basic nitrogen of histidine is used to abstract aproton from serine, threonine or cysteine to activate it as anucleophile. In a histidine proton shuttle, histidine is used to quicklyshuttle protons, it can do this by abstracting a proton with its basicnitrogen to make a positively-charged intermediate and then use anothermolecule, a buffer, to extract the proton from its acidic nitrogen. Incarbonic anhydrases, a histidine proton shuttle is utilized to rapidlyshuttle protons away from a zinc-bound water molecule to quicklyregenerate the active form of the enzyme. The amino acid is a precursorfor histamine and carnosine biosynthesis.

Histidine has two enantiomeric forms: D-histidine and L-histidine. Thestructure of histidine is shown below. Histidine is a basic, essentialamino acid that is also a precursor of histamine, a compound released byimmune system cells during an allergic reaction. Histamine is needed forgrowth and for the repair of tissue, as well as the maintenance of themyelin sheaths that act as protector for nerve cells. It is furtherrequired for the manufacture of both red and white blood cells, andhelps to protect the body from damage caused by radiation and inremoving heavy metals from the body. In the stomach, histidine is alsohelpful in producing gastric juices, and people with a shortage ofgastric juices or suffering from indigestion, may also benefit from thisnutrient. Histidine is also used for sexual arousal, functioning andenjoyment. Histidinemia is an inborn error of the metabolism ofhistidine due to a deficiency of the enzyme histidase, where high levelsof histidine are found in the blood and urine, and may manifest inspeech disorders and mental retardation.

The compositions described herein that comprise chromium and histidine,or chromium histidinate complexes, such as chromium histidinate chromiumtrihistidinate, and chromium polyhistidinate, or combinations thereof,e.g., including components of some of the compositions disclosed hereinfor the administration of chromium over a period of time, exhibitimproved absorption in mammals over other known chromium complexes. Inparticular, the compositions described herein show superior absorptionand intracellular release of chromium from the histidinate complex.

Several of the compositions disclosed herein can include chromium andhistidine, or chromium histidinate complexes in combination with otherchromium complexes including chromium picolinate, chromium nicotinate,chromium chloride, tri-chromium(III) oxo acetate cluster([Cr(3)O(OAc)(6)](+)), biomimetic cation[Cr(3)O(O(2)CCH(2)CH(3))(6)(H(2)O)(3)](+) and chromium triphenylanine,and any other chromium complex now known or discovered in the future.

The compositions described herein can contain one or more chiral centersand/or double bonds and, therefore, exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers, ordiastereomers. The chemical structures depicted herein, and thereforethe compositions disclosed herein, encompass all of the correspondingcompounds' or compositions' enantiomers and stereoisomers, that is, boththe stereomerically pure form (e.g., geometrically pure,enantiomerically pure, or diastereomerically pure) and enantiomeric andstereoisomeric mixtures.

As used herein, a composition that “substantially” comprises a compoundmeans that the composition contains more than about 80% by weight, morepreferably more than about 90% by weight, even more preferably more thanabout 95% by weight, and most preferably more than about 97% by weightof the compound. As used herein, a composition that “substantially”comprises a chromium complex refers to a composition that contains morethan or equal to 7.0% of trivalent or dietary chromium. Preferably, acertificate of analysis for the compositions disclosed herein indicatethat the compositions are negative for microbial growth, yeast and moldshould be present in less than 300 cells/g and the toxic metals shouldbe less than 1 ppm.

In some embodiments, the compositions disclosed herein are in the formof pharmaceutically effective salts. The phrase “pharmaceuticallyacceptable salt(s),” as used herein includes, but is not limited to,salts of acidic or basic groups that may be present in the compositionsdisclosed herein. Compounds that are basic in nature are capable offorming a wide variety of salts with various inorganic and organicacids. The acids that may be used to prepare pharmaceutically acceptableacid addition salts of such basic compounds are those that formnon-toxic acid addition salts, i.e., salts containing pharmacologicallyacceptable anions, including but not limited to sulfuric, citric,maleic, acetic, oxalic, hydrochloride, hydrobromide, hydroiodide,nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate,acetate, lactate, salicylate, citrate, acid citrate, tartrate, oleate,tannate, pantothenate, bitartrate, ascorbate, succinate, maleate,gentisinate, fumarate, gluconate, glucaronate, saccharate, formate,benzoate, glutamate, methanesulfonate, ethanesulfonate,benzenesulfonate, p-toluenesulfonate and pamoate (i.e.,1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Compounds present inthe compositions disclosed herein that include an amino moiety also canform pharmaceutically acceptable salts with various amino acids, inaddition to the acids mentioned above. Compounds present in thecompositions disclosed herein that are acidic in nature are capable offorming base salts with various pharmacologically acceptable cations.Non limiting examples of such salts include alkali metal or alkalineearth metal salts and, particularly, calcium, magnesium, sodium lithium,zinc, potassium, silicon, phosphorus and iron salts.

As used herein, the term “hydrate” means a compound or a salt thereof,that further includes a stoichiometric or non-stoichiometric amount ofwater bound by non-covalent intermolecular forces. The term hydrateincludes solvates, which are stoichiometric or non-stoichiometricamounts of a solvent bound by non-covalent intermolecular forces.Preferred solvents are volatile, non-toxic, and/or acceptable foradministration to humans in trace amounts.

The amount of a compound of the invention that will be effective in thetreatment of a particular disorder or condition disclosed herein willdepend on the nature of the disorder or condition, and can be determinedby standard clinical techniques. In addition, in vitro or in vivo assaysmay optionally be employed to help identify optimal dosage ranges. Theprecise dose to be employed in the compositions will also depend on theroute of administration, and the seriousness of the disease or disorder,and should be decided according to the judgment of the practitioner andeach circumstances. However, suitable dosage ranges for oraladministration are generally about 0.001 milligram to 5000 milligrams ofa total chromium complex (e.g., fast-acting and slow-acting combined)per kilogram body weight. In preferred embodiments, the oral dose is0.01 milligram total chromium complex to 1000 milligrams per kilogrambody weight, more preferably 0.1 milligram to 100 milligrams perkilogram body weight, more preferably 0.5 milligram to 25 milligrams perkilogram body weight, and yet more preferably 1 milligram to 10milligrams per kilogram body weight. The dosage amounts described hereinrefer to total amounts administered; that is, if more than one chromiumcomplex or more than one composition disclosed herein is administered,the preferred dosages correspond to the total amount of the compositionsdisclosed herein administered. Oral compositions preferably contain 10%to 95% active ingredient.

The compositions disclosed herein can preferably be formulated withother active ingredients as a slow-acting agent or long acting agent inaddition to drugs or alone before meals and/or after meals. Effectivedoses may be extrapolated from dose-response curves derived from invitro or animal model test systems. Such animal models and systems arewell known in the art.

In accordance with the methods disclosed herein, the amount of chromiumprovided by the compositions that comprise fast-acting and slow-actingcomplexes disclosed herein can be at least 50 μg per day, for example atleast 60 μg, at least 70 μg, at least 80 μg, at least 90 μg, at least100 μg, at least 124 μg, at least 150 μg, at least 200 μg, at least 250μg, at least 300 μg, at least 350 μg, at least 400 μg, at least 450 μg,at least 500 μg, at least 550 μg, at least 600 μg, at least 650 μg, atleast 700 μg, at least 750 μg, at least 800 μg, at least 850 μg, atleast 900 μg, at least 950 μg, at least 1,000 μg, at least 1500 μg, atleast 2,000 μg, at least 2500 μg, at least 3000 μg, at least 3500 μg, atleast 4000 μg, at least 4500 μg or at least 5000 μg chromiumcomplex/day.

The fast-acting and the slow-acting chromium complexes can be providedto a subject such that the ratio of chromium in the form of a“fast-acting” chromium complex to the chromium in the form of a“slow-acting” chromium complex is anywhere from 10:1 to 1:10, e.g., 9:1,8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1;8, 1:9, 1:10, or any fraction in between. In some embodiments, the ratioof chromium provided in the form of a fast-acting chromium complex tothe slow-acting chromium complex is 1:1.

By way of example, the level of chromium used for supplementation inorder to inhibit the onset of insulin resistance is at least about 50μg/day. Chromium picolinate and chromium chloride have been administeredto rats at levels several thousand times the upper limit of theestimated safe and adequate daily dietary intake (ESADDI) for chromiumfor humans (based on body weight) without toxic effects. R. Anderson etal., Lack of Toxicity of Chromium Chloride and Picolinate, 16 J. Am.Coll. Nutr. 273-279 (1997). While the level of chromium, in the form offast-acting and slow-acting chromium complexes, used for supplementationmay be within several thousand times the upper limit of the ESADDI,preferably, the total amount of chromium provided by the fast-acting andslow-acting complexes is between about 50 and 2,000 μg/day. Morepreferably, the amount of total chromium provided by the fast-acting andslow-acting complexes is between about 300 and 1,000 μg/day. Mostpreferably, the amount of total chromium is between about 400 and 1,000μg/day. In a particularly preferred embodiment, the amount of totalchromium is between about 600 and 1,000 μg/day. These doses are based ona 70 kg adult human, and that the dose can be applied on a per-kilogrambasis to humans or animals of different weights.

In some embodiments, the amount of fast-acting chromium complex and theamount of slow-acting chromium complex in a composition provide agreater than additive effect in lowering serum glucose levels thaneither complex alone. In some embodiments, the amount of fast-actingchromium complex and the amount of slow-acting chromium complex in acomposition provide a synergistic or greater than additive effect inimproving insulin sensitivity. In some embodiments, the amount offast-acting chromium complex and the amount of slow-acting chromiumcomplex in a composition provide a greater than additive effect intreating dyslipidemia. In some embodiments, the amount of fast-actingchromium complex and the amount of slow-acting chromium complex in acomposition provide a greater than additive effect in lowering serumcholesterol, lowering serum triglycerides, lowering free fatty acidlevels, or increasing HDL levels. In some embodiments, the amount offast-acting chromium complex and the amount of slow-acting chromiumcomplex in a composition provide a greater than additive effect inincreasing lean muscle mass.

In some embodiments, the compositions disclosed herein are provided inan amount effective for the prevention of insulin resistance. As usedherein, the term “insulin resistance (IR)” refers to a physiologicallyabnormal state in which cells do not respond appropriately to insulin,such that glucose in the blood cannot efficiently enter cells and,therefore, leads to hyperglycemia. Insulin resistance is believed toaffect one in three adult Americans which amounts to approximately 99.3million Americans with some degree of insulin resistance. Thecardiovascular and metabolic disturbances associated with IR canindividually and interdependently lead to a substantial increase incardiovascular disease (CVD) morbidity and mortality, making thecardiometabolic syndrome an established and strong risk factor forpremature and severe CVD and stroke. For example, in some embodiments, asubject is provided a composition comprising a fast-acting chromiumcomplex chromium such as histidinate in combination with a sufficientamount of a slow-acting chromium complex to inhibit or reduce the riskof the onset of insulin resistance are provided. The fast-actingchromium complex can be formulated together with the slow-actingchromium complex or the fast-acting chromium complex can be administeredseparately before, substantially contemporaneously with or after theslow-acting chromium complex. The assessment of the affects of thecompositions on insulin resistance can readily be determined usingroutine techniques known to those skilled in the art, and described, forexample, in U.S. patent application Ser. No. 10/090,038, hereinincorporated by reference in its entirety. Preferably, the sufficientamount of chromium provided by the fast-acting and slow-acting chromiumcomplexes and contained in the composition is between about 50 μg and2000 μg.

Advantageously, an individual is administered a pharmaceuticallyeffective dose of a hydrophilic chromium complex such as chromiumhistidinate in combination with at least one other lipophilic chromiumcomplex. In some embodiments, a composition the fast-acting and aslow-acting chromium complexes are administered substantiallysimultaneously. In an alternative embodiment, the fast-acting,hydrophilic and slow-acting, lipohilic chromium complexes are providedto the subject sequentially in either order. If administered separately,the fast-acting and slow-acting chromium complex should be given in atemporally proximate manner, e.g., within a twenty-four hour period.More particularly, the a fast-acting and a slow-acting chromium complex)can be given within one hour of each other. One of skill in the art willappreciate that other components may be added separately or incorporatedinto a single formulation to enhance the effects of chromium.

In some embodiments, the compositions disclosed herein can be providedprior to or concomitantly with an insulin resistance-inducing food.Insulin resistance-inducing foods generally have high glycemic indexes,e.g., over 50. In other embodiments, the compositions are provided afterthe insulin resistance inducing food. In embodiments wherein thecompositions and the insulin resistance-inducing foods are not providedconcomitantly, the composition and the food are preferably provided in atemporally proximate manner, e.g., within twenty four hours, and morepreferably within one hour.

In some embodiments, uncomplexed chelating agents are advantageouslyincluded in the compositions to facilitate absorption of other ingestedchromium as well as other metals including, but not limited to, copper,iron, magnesium, manganese, and zinc. Suitable chelating agents includehistidine, any essential amino D or L amino acids, tri amino acidformulae including but not limited to, triphenylalanine, trihistidine,triarginine, picolinic acid, nicotinic acid, or both picolinic acid andnicotinic acid. Thus, the compositions of the disclosed invention arereadily absorbable forms of chromium complex which also facilitateabsorption of other essential metals in the human diet. In someembodiments, certain chelating agents may be added to facilitateabsorption of the chromium complex, or combination of chromium complexesin the compositions disclosed herein. Chelating agents such ashistidine, picolinic acid and nicotinic acid are available from manycommercial sources, including Sigma-Aldrich (St. Louis, Mo.) (picolinicacid; catalog No. P5503; nicotinic acid; catalog No. PN4126).Preferably, the ratio of either the fast-acting, or slow-acting, or thecombination of the fast-acting and slow-acting chromium complex to thechelating agent from about 10:1 to about 1:10 (w/w), more preferablyfrom about 5:1 to about 1:5 (w/w). Alternatively, the molar ratio ofchromium complex to the uncomplexed chelating agent is preferably 1:1,and may be from about 5:1 to about 1:10. The chelating agents with D orL amino acid and or with tri or mono and di forms of chromium complexwith tri amino acid or one or more amino acids but not limited tochromium triphenylanine, chromium trihistidine, chromiumpolyphenylanine, chromium poly hisitidine, chromium polynicotinate,chromium diphenylananine, chromium dipicolinic acid, chromiumdihisitidine etc. More than one chelating agent, e.g. both nicotinic andpicolinic acid can be included in the compositions disclosed herein, oradministered to subject in the methods described herein.

The administration of the compositions disclosed herein can be by any ofthe methods of administration described below or by delivery methodsknown by one of skill in the art. The compositions may be administeredorally, through parenteral nutrition, e.g., feeding tube orintravenously, and through other known means. Chromium histidinate incombination with other chromium complexes or essential nutrients but notlimited to fatty acids, carbohydrates, minerals and vitamins etc. is aparticularly preferred source fast-acting chromium complex due to itshigh level of bioavailability, but other fast-acting, hydrophilicchromium complex can also be used.

For oral administration, the compositions disclosed herein can beprovided as a tablet, aqueous or oil suspension, dispersible powder orgranule, emulsion, hard or soft capsule, syrup, elixir, or beverage.Compositions intended for oral use can be prepared according to anymethod known in the art for the manufacture of pharmaceuticallyacceptable compositions and such compositions may contain one or more ofthe following agents: sweeteners, flavoring agents, coloring agents andpreservatives. The sweetening and flavoring agents will increase thepalatability of the preparation. Tablets containing chromium complexesin admixture with non-toxic pharmaceutically acceptable excipientssuitable for tablet manufacture are acceptable. Pharmaceuticallyacceptable vehicles such as excipients are compatible with the otheringredients of the formulation (as well as non-injurious to thepatient). Such excipients include inert diluents such as calciumcarbonate, sodium carbonate, lactose, calcium phosphate or sodiumphosphate; granulating and disintegrating agents, such as corn starch oralginic acid; binding agents such as starch, gelatin or acacia; andlubricating agents such as magnesium stearate, stearic acid or talc.Tablets can be uncoated or can be coated by known techniques to delaydisintegration and absorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period of time. For example, atime delay material such as glyceryl monostearate or glyceryl distearatealone or with a wax can be employed.

Formulations for oral use can also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, such as peanut oil, liquid paraffin or olive oil. Aqueoussuspensions can contain the chromium complex of the invention inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include suspending agents, dispersing orwetting agents, one or more preservatives, one or more coloring agents,one or more flavoring agents and one or more sweetening agents such assucrose or saccharin.

Oil suspensions can be formulated by suspending the active ingredient ina vegetable oil, such as arachis oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oil suspension cancontain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents can be added to provide a palatable oral preparation. Thesecompositions can be preserved by an added antioxidant such as ascorbicacid. Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Additional excipients,for example sweetening, flavoring and coloring agents, can also bepresent.

Syrups and elixirs can be formulated with sweetening agents, such asglycerol, sorbitol or sucrose. Such formulations can also contain ademulcent, a preservative, a flavoring or a coloring agent.

The chromium complex preparations for parenteral administration can bein the form of a sterile injectable preparation, such as a sterileinjectable aqueous or oleaginous suspension. This suspension can beformulated according to methods well known in the art using suitabledispersing or wetting agents and suspending agents. The sterileinjectable preparation can also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,such as a solution in 1,3-butanediol. Suitable diluents include, forexample, water, Ringer's solution and isotonic sodium chloride solution.In addition, sterile fixed oils can be employed conventionally as asolvent or suspending medium. For this purpose, any bland fixed oil canbe employed including synthetic mono or diglycerides. In addition, fattyacids such as oleic acid can likewise be used in the preparation ofinjectable preparations.

The pharmaceutical compositions can also be in the form of oil-in-wateremulsions. The oily phase can be a vegetable oil, such as olive oil orarachis oil, a mineral oil such as liquid paraffin, or a mixturethereof. Suitable emulsifying agents include naturally-occurring gumssuch as gum acacia and gum tragacanth, naturally occurring phosphatides,such as soybean lecithin, esters or partial esters derived from fattyacids and hexitol anhydrides, such as sorbitan mono-oleate, andcondensation products of these partial esters with ethylene oxide, suchas polyoxyethylene sorbitan mono-oleate. The emulsions can also containsweetening and flavoring agents.

It will be appreciated by the skilled artisan that the amount ofchromium histidine in combination with chromium complex that can becombined with the carrier material to produce a single dosage form willvary depending upon the host treated and the particular mode ofadministration.

When administered to a mammal, e.g., to an animal for veterinary use orfor improvement of livestock, or to a human for therapeutic use, thecompositions disclosed herein are administered in isolated form or asthe isolated form in a therapeutic composition. As used herein,“isolated” means that the compositions disclosed herein are separatedfrom other components of either (a) a natural source, such as a plant orcell or food, preferably bacterial culture, or (b) a synthetic organicchemical reaction mixture. Preferably, via conventional techniques, thecompositions disclosed herein are purified. As used herein, “purified”means that when isolated, the isolate contains at least 95%, preferablyat least 98% of the composition.

In some embodiments, the compositions disclosed herein are provided tothe subject orally. In other embodiments, the compositions disclosedherein are provided by any other convenient route, for example, byintravenous infusion or bolus injection, by absorption throughepithelial or mucocutaneous linings (e.g., oral mucosa, rectal andintestinal mucosa, etc.) and can be administered together with anotherbiologically active agent. Administration can be systemic or local.Various delivery systems useful in the methods disclosed herein includefor example, encapsulation in liposomes, microparticles, microcapsules,capsules, etc., and can be used to administer a compound of theinvention. In certain embodiments, more than one composition disclosedherein is administered to an individual.

Other modes of administration useful in the methods include but are notlimited to intradermal, intramuscular, intraperitoneal, intravenous,subcutaneous, intranasal, epidural, oral, sublingual, intranasal,intracerebral, intravaginal, transdermal, rectally, by inhalation, ortopically, particularly to the ears, nose, eyes, or skin. The preferredmode of administration is left to the discretion of the professional,and will depend in-part upon the site of the condition to be treated. Inmost instances, administration will result in the release of thecompositions disclosed herein into the bloodstream.

In specific embodiments, it can be desirable to administer one or morecompositions disclosed herein locally to the area in need of treatment.This can be achieved, for example, and not by way of limitation, bylocal infusion during surgery, topical application, e.g., in conjunctionwith a wound dressing after surgery, by injection, by means of acatheter, by means of a suppository, or by means of an implant, saidimplant being of a porous, non-porous, or gelatinous material, includingmembranes, such as sialastic membranes, or fibers. In one embodiment,administration can be by direct injection at the site (or former site)of an atherosclerotic plaque tissue

In certain embodiments, for example, for the treatment of Alzheimer'sdisease, it may be desirable to introduce one or more compositionsdisclosed herein into the central nervous system by any suitable route,including intraventricular, intrathecal or epidural injection.Intraventricular injection may be facilitated by an intraventricularcatheter, for example, attached to a reservoir, such as an Ommayareservoir.

Pulmonary administration can also be employed, e.g., by use of aninhaler or nebulizer, and formulation with an aerosolizing agent, or viaperfusion in a fluorocarbon or synthetic pulmonary surfactant. Incertain embodiments, the compositions disclosed herein can be formulatedas a suppository, with traditional binders and vehicles such astriglycerides.

Preferably, the compositions disclosed herein are formulated with apharmaceutically acceptable vehicle. As used herein, the term“pharmaceutically acceptable” means approved by a regulatory agency ofthe Federal or a state government or listed in the U.S. Pharmacopeia orother generally recognized pharmacopeia for use in animals, and moreparticularly in humans. The term “vehicle” refers to a diluent,adjuvant, excipient, or carrier with which a compound of the inventionis administered. Such pharmaceutical vehicles can be liquids, such aswater and oils, including those of petroleum, animal, vegetable orsynthetic origin, such as peanut oil, soybean oil, mineral oil, sesameoil and the like. The pharmaceutical vehicles can be saline, gum acacia,gelatin, starch paste, talc, keratin, colloidal silica, urea, and thelike. In addition, auxiliary, stabilizing, thickening, lubricating andcoloring agents may be used. When administered to a patient, thecompositions of the invention and pharmaceutically acceptable vehiclesare preferably sterile. Water is a preferred vehicle when thecompositions of the invention is administered intravenously. Salinesolutions and aqueous dextrose and glycerol solutions can also beemployed as liquid vehicles, particularly for injectable solutions.Suitable pharmaceutical vehicles also include excipients such as starch,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, sodium stearate, glycerol monostearate, talc, sodium chloride,dried skim milk, glycerol, propylene, glycol, water, ethanol and thelike. The present compositions, if desired, can also contain minoramounts of wetting or emulsifying agents, or pH buffering agents.

The present compositions can take the form of solutions, suspensions,emulsion, tablets, pills, pellets, capsules, capsules containingliquids, powders, sustained-release formulations, suppositories,emulsions, aerosols, sprays, suspensions, or any other form suitable foruse.

In some embodiments, the compositions disclosed herein are formulatedfor oral delivery, for example in the form of tablets, lozenges, aqueousor oily suspensions, granules, powders, emulsions, capsules, syrups, orelixirs. Compounds and compositions described herein for oral deliverycan also be formulated in foods and food mixes. Orally administeredcompositions can contain one or more optionally agents, for example,sweetening agents such as fructose, aspartame or saccharin; flavoringagents such as peppermint, oil of wintergreen, or cherry; coloringagents; and preserving agents, to provide a pharmaceutically palatablepreparation. Moreover, where in tablet or pill form, the compositionscan be coated to delay disintegration and absorption in thegastrointestinal tract thereby providing a sustained action over anextended period of time. Selectively permeable membranes surrounding anosmotically active driving compound are also suitable for orallyadministered compounds and compositions described herein. In these laterplatforms, fluid from the environment surrounding the capsule is imbibedby the driving compound, which swells to displace the agent or agentcomposition through an aperture. These delivery platforms can provide anessentially zero order delivery profile as opposed to the spikedprofiles of immediate release formulations. A time delay material suchas glycerol monostearate or glycerol stearate can also be used. Oralcompositions can include standard vehicles such as mannitol, lactose,starch, magnesium stearate, sodium saccharine, cellulose, magnesiumcarbonate, etc. Such vehicles are preferably of pharmaceutical grade.

In some embodiments, the compositions described herein can be in theform of nutraceutical packs not limited to functional foods, beverages,bars, dietary supplements, capsules, powder form or gelatin form,pharmaceutical packs or kits comprising one or more containers filledwith one or more compositions disclosed herein. Optionally associatedwith such container(s) can be a notice in the form prescribed by agovernmental agency regulating the manufacture, use or sale ofpharmaceuticals or biological products, which notice reflects approvalby the agency of manufacture, use or sale for human administration. In acertain embodiment, the kit contains more than one compound describedherein. In another embodiment, the kit comprises a compound describedherein and another lipid-mediating compound, glycemic control andantihypertensive drugs, including but not limited to insulin, statin, athiazolidinedione, or a fibrate or dietary modifications.

The compositions disclosed herein can be assayed in vitro and in vivo,for the desired therapeutic or prophylactic activity, prior to use inhumans. For example, in vitro assays can be used to determine whetheradministration of a specific compound described herein or a combinationof compositions disclosed herein is preferred for lowering fatty acidsynthesis. The compositions disclosed herein also be demonstrated to beeffective and safe using animal model systems.

Throughout the specification there are references to identifying asubject in need of administration of a chromium complex which includes ahydrophilic chromium complex and a lipophilic chromium complex. The termidentification is not intended to be limiting and includes in eachinstance a belief by the subject that the chromium complexes willbenefit the subject, self-identification, and identification by thirdparty using various techniques. The identification may be of at leastone condition selected from the group consisting of: insulin resistance,cardiovascular disease, progressive renal disease, end stage renaldisease, endothelial dysfunction, left ventricular hypertrophy, cardiachyperreactivity, dyslipidemia, hyperglycemia, enhanced renninangiotensin activity, aldosterone syndrome, impaired pressurenatriuresis, chronic low grade inflammation, diabetes mellitus,hypertension, atherosclerosis, micoralbuminuria, obesity, depression,Syndrome X, and polycystic ovary syndrome. The identification maycomprise identifying an individual that is taking a compositioncomprising a compound selected from the group consisting of:non-steroidal anti inflammatory compounds, oral contraceptives,implantable steroid contraceptives, hormone replacement therapy, betablockers, potassium channel openers and immunosuppressive drugs.

Therapeutic Uses of Chromium Histidine/Histidinate

In accordance with the methods disclosed herein, a compositioncomprising, consisting essentially of, or consisting of a chromium andhistidine, chromium histidinate complex, chromium trihistidinate, orchromium polyhistidinate complex, or any combination thereof, and asecond chromium complex comprising, consisting essentially of orconsisting of a slow-acting chromium complex such as chromium picolinateis provided to a subject, such as a mammal, with or at risk of aging,Alzheimer's Disease, cancer, cardiovascular disease, diabeticnephropathy, diabetic retinopathy, a disorder of glucose metabolism,dyslipidemia, dyslipoproteinemia, enhancing bile production, enhancingreverse lipid transport, hypertension, impotence, inflammation, insulinresistance, lipid elimination in bile, modulating C reactive protein,obesity, oxysterol elimination in bile, pancreatitis, Parkinson'sdisease, a peroxisome proliferator activated receptor-associateddisorder, phospholipid elimination in bile, renal disease, septicemia,metabolic syndrome disorders (e.g., Syndrome X), a thrombotic disorder,gastrointestinal disease, irritable bowel syndrome (IBS), inflammatorybowel disease (e.g., Crohn's Disease, ulcerative colitis), arthritis(e.g., rheumatoid arthritis, osteoarthritis), autoimmune disease (e.g.,systemic lupus erythematosus), scleroderma, ankylosing spondylitis, goutand pseudogout, muscle pain: polymyositis/polymyalgiarheumatica/fibrositis; infection and arthritis, juvenile rheumatoidarthritis, tendonitis, bursitis and/or other soft tissue rheumatism.

As used herein, the term “treatment” or “treating” refers to anamelioration of a disease or disorder, or at least one discerniblesymptom thereof. The term “treatment” or “treating” refers to inhibitingthe progression of a disease or disorder, either physically, e.g.,stabilization of a discernible symptom, or physiologically, e.g.,stabilization of a physical parameter, or both.

In certain embodiments, the compositions disclosed herein are providedto a subject, such as a mammal, as a preventative measure against suchdiseases. As used herein, “prevention” or “preventing” refers to areduction of the risk of acquiring a given disease or disorder alone orin combination with other clinical condition.

In some embodiments, the compositions disclosed herein are provided as apreventative measure to a patient, preferably a human having a geneticpredisposition to a aging, Alzheimer's Disease, cancer, cardiovasculardisease, diabetic nephropathy, diabetic retinopathy, a disorder ofglucose metabolism, dyslipidemia, dyslipoproteinemia, enhancing bileproduction, enhancing reverse lipid transport, hypertension, impotence,inflammation, insulin resistance, lipid elimination in bile, modulatingC-reactive protein, obesity, oxysterol elimination in bile,pancreatitis, Parkinson's disease, a peroxisome proliferator activatedreceptor-associated disorder, phospholipid elimination in bile, renaldisease, septicemia, metabolic syndrome disorders (e.g., Syndrome X), athrombotic disorder, inflammatory processes and diseases likegastrointestinal disease, irritable bowel syndrome (IBS), inflammatorybowel disease (e.g., Crohn's Disease, ulcerative colitis), arthritis(e.g., rheumatoid arthritis, osteoarthritis), autoimmune disease (e.g.,systemic lupus erythematosus), scleroderma, ankylosing spondylitis, goutand pseudogout, muscle pain: polymyositis/polymyalgiarheumatica/fibrositis; infection and arthritis, juvenile rheumatoidarthritis, tendonitis, bursitis and other soft tissue rheumatism.Examples of such genetic predispositions include but are not limited tothe di-electcons 4 allele of apolipoprotein E, which increases thelikelihood of Alzheimer's Disease; a loss of function or null mutationin the lipoprotein lipase gene coding region or promoter (e.g.,mutations in the coding regions resulting in the substitutions D9N andN291S; for a review of genetic mutations in the lipoprotein lipase genethat increase the risk of cardiovascular diseases, dyslipidemias anddyslipoproteinemias (Hayden and Ma, 1992, Mol. Cell Biochem. 113:171176); and familial combined hyperlipidemia and familialhypercholesterolemia.

In some embodiments, the compositions disclosed herein are provided to asubject as a preventative measure to a mammal having a non-geneticpredisposition to cardiometabolic syndrome, in aging, Alzheimer'sDisease, cancer, cardiovascular disease, diabetic nephropathy, diabeticretinopathy, a disorder of glucose metabolism, dyslipidemia,dyslipoproteinemia, enhancing bile production, enhancing reverse lipidtransport, hypertension, impotence, inflammation, insulin resistance,lipid elimination in bile, modulating C-reactive protein, obesity,oxysterol elimination in bile, pancreatitis, Parkinson's disease, aperoxisome proliferator activated receptor-associated disorder,phospholipid elimination in bile, renal disease, septicemia, metabolicsyndrome disorders (e.g., Syndrome X), a thrombotic disorder,inflammatory processes and diseases like gastrointestinal disease,irritable bowel syndrome (IBS), inflammatory bowel disease (e.g.,Crohn's Disease, ulcerative colitis), arthritis (e.g., rheumatoidarthritis, osteoarthritis), autoimmune disease (e.g., systemic lupuserythematosus), scleroderma, ankylosing spondylitis, gout andpseudogout, muscle pain: polymyositis/polymyalgia rheumatica/fibrositis;infection and arthritis, juvenile rheumatoid arthritis, tendonitis,bursitis and other soft tissue rheumatism. Examples of such non-geneticpredispositions include but are not limited to cardiac bypass surgeryand percutaneous transluminal coronary angioplasty, which often lead torestenosis, an accelerated form of atherosclerosis; diabetes in women,which often leads to polycystic ovarian disease; and cardiovasculardisease, which often leads to impotence. Accordingly, the compositionsdescribed herein can be used for the prevention of one disease ordisorder and concurrently treating another disease (e.g., prevention ofpolycystic ovarian disease while treating diabetes; prevention ofimpotence while treating a cardiovascular disease).

In some embodiments, the compositions disclosed herein are provided to asubject to inhibit the onset of insulin resistance in a subject based oncriteria including but not limited to family history, diet and drug use.In some embodiments, for example, an individual at risk for developinginsulin resistance may believe it is in need or may self-identify or beidentified based on family history, obesity, diabetes, CVD and otherassociated disease conditions including depression, mental healthdiseases or disorders, glucose and lipid metabolism disturbances, a diethigh in fats, carbohydrates, low dietary fiber, deficiency of essentialnutrients and not limited to drugs such as a statin drug, non-steroidalanti-inflammatory drug, steroid, oral contraceptive, hormone replacementtherapy drug, beta blocker, potassium channel opener, or diuretic anddepression drugs. Accordingly, some embodiments provide a method forinhibiting the development of drug-induced insulin resistance includingadministering a fast-acting, hydrophilic and a slow-acting, lipophilicchromium complex to an individual receiving a contemporaneous dose of adrug that induces insulin resistance. Advantageously, the amount ofchromium complex administered is an amount effective to inhibit thedevelopment of insulin resistance.

The administration of an effective dose of a composition describedherein (e.g., chromium histidinate and chromium picolinate), to subjectswho have a diet or take drugs which have been linked with the onset ofinsulin resistance actually inhibits or attenuates the onset of insulinresistance. The supplementation with a fast-acting chromium andslow-acting complex, inhibits the induction of insulin resistance. Bynot developing insulin resistance in the first place, the patient is notexposed to the associated diseases and risks. The patient also does notneed to take additional, and sometimes costly, medications to treat theinsulin resistance and associated diseases.

Without being limited to a particular theory, we propose thatcombination chromium supplementation with fast and slow-acting chromiumcomplex inhibits insulin resistance from developing by improving insulinsensitivity and lipid profile levels and lowering blood sugar.Accordingly, in one embodiment, a method of inhibiting or reducing therisk of insulin resistance through combination chromium supplementationis provided.

Chromium supplementation includes the administration of a fast-actingchromium complex such as histidinate in combination with at least oneother slow-acting chromium complex to an individual. Advantageously, thefast and slow-acting chromium complexes are synthetic. The synthesis anduse of chromium picolinates, for example, is described in U.S. Pat. Nos.Re 33,988 and 5,087,623, the entire contents of which are herebyincorporated herein by reference in their entirety. Chromictripicolinate is available from health food stores, drug stores andother commercial sources. The synthesis and use of chromicpolynicotinate is described in U.S. Pat. No. 5,194,615.

Inhibition of insulin resistance is accomplished by administering aneffective dose of fast-acting chromium complex such as a chromiumhistidinate complex with a slow-acting chromium complex such as chromiumpicolinate to an individual separately or as a single composition. Asubject can begin chromium supplementation at the beginning of theirtreatment with insulin-resistance. Alternatively, the subject can beginsupplementation with a chromium complex after the subject's treatmentwith insulin-resistance has begun, but before developing insulinresistance.

Insulin resistance is a key pathogenic parameter of Type 2 diabetes, andclinical interventions that improve insulin sensitivity are consideredcornerstones in the management of the disease. In addition, therelationship of insulin resistance to cardiovascular disease and itsassociated risk factors has been well established over the past fewyears. Therefore, in a preferred embodiment, methods and compositionsfor thwarting the development of insulin resistance are providedcomprising the administration of a fast-acting and slow-acting chromiumcomplexes with a hypoglycemic drug such as metformin, which inhibitsinsulin resistance from developing. Combinations of pharmacologic agents(such as sulfonylureas/metformin, sulfonylureas/glitazones, andmetformin/glitazones) are highly effective pharmacologic interventionsthat appear to lower both glucose and insulin levels. Further, there isevidence that triple drug therapy (e.g.sulfonylureas/metformin/glitazones) can lower clinical glycemia inaddition to lowering insulin levels. Hence, in some embodiments,compositions comprising a hydrophobic chromium complex and a hydrophilicchromium complex with metformin, sulfonylureas, and glitazones orcombinations thereof are administered to a subject taking drugs whichinduce insulin resistance to inhibit the onset of such insulinresistance.

The disclosure represents the present technology in that the subject hasa chance of developing insulin resistance or diabetes or associatedconditions but not limited to cardiovascular disease, obesity, diabetes,combination one or two disease conditions based on ATPIII guidelines andor due to mental health conditions such as depression, schizophrenia,alzheimers disease and other conditions such HIV and HIV lipodystrophyand polycystic ovary syndrome. The insulin resistance might be due tofamily history, body weight, diet and drugs.

Treatment of Cardiovascular Diseases

The present invention provides methods for the treatment or preventionof a cardiovascular disease, comprising administering to a patient atherapeutically effective amount of a composition comprising, consistingessentially of, or consisting of chromium and histidine, or a chromiumhistidinate complex with a slow-acting chromium and a pharmaceuticallyacceptable vehicle. As used herein, the term “cardiovascular diseases”refers to diseases of the heart and circulatory system. These diseasesare often associated with dyslipoproteinemias and/or dyslipidemiasand/or familial hyperlipoproteinemias, hyper cholesterolemia and hyperlipidemia. Cardiovascular diseases which the compositions of the presentinvention are useful for preventing or treating include but are notlimited to arteriosclerosis; atherosclerosis; stroke; ischemia;endothelium dysfunctions, in particular those dysfunctions affectingblood vessel elasticity; peripheral vascular disease; coronary heartdisease; myocardial infarction; cerebral infarction and restenosis.

The compositions disclosed herein, e.g., a fast-acting chromium such aschromium histidinate with a slow-acting chromium complexes, e.g.chromium picolinate, are preferably used in methods for treatingcardiovascular disease and its related pathologies, including, forexample, hypertrophy, hypertension, congestive heart failure, myocardialischemia, ischemia reperfusion injuries in an organ, arrhythmia, andmyocardial infarction. One embodiment is directed to a method oftreating cardiovascular disease in a mammal by concurrentlyadministering to the mammal a therapeutically effective amount of atherapeutic cardiovascular compound, and a hydrophilic chromium complexand a lipophilic chromium complex. Therapeutic cardiovascular compoundssuitable for use in methods described herein include an angiotensinconverting enzyme inhibitor, an angiotensin II receptor antagonist, acalcium channel blocker, an anti-thrombotic agent, a β-adrenergicreceptor antagonist, a vasodilator, a diuretic, an α-adrenergic receptorantagonist, an antioxidant, and a mixture thereof. In some embodiments,the therapeutic cardiovascular compound is PPADS.

Treatment of Dyslipidemias

Also provided are methods for the treatment or prevention of adyslipidemia comprising administering to a patient a therapeuticallyeffective amount of composition disclosed herein, i.e., a fast-actinghydrophilic chromium complex in combination with a slow-acting orlipophilic chromium complex, e.g., chromium histidinate complex, achromium picolinate complex, and a pharmaceutically acceptable vehicle.

As used herein, the term “dyslipidemias” refers to disorders that leadto or are manifested by aberrant levels of circulating lipids. To theextent that levels of lipids in the blood are too high, the compositionsdescribed herein are administered to a patient to restore normal levels.Normal levels of lipids are reported in medical treatises known to thoseof skill in the art. For example, recommended blood levels of LDL, HDL,free triglycerides and others parameters relating to lipid metabolismcan be found at the web site of the American Heart Association and thatof the National Cholesterol Education Program of the National Heart,Lung and Blood Institute(http://www.americanheart.org/cholesterol/about_level.html andhttp://www.nhlbi.nih.gov/health/public/heart/chol/hbc_what.html,respectively). At the present time, the recommended level of HDLcholesterol in the blood is above 35 mg/dL; the recommended level of LDLcholesterol in the blood is below 70 mg/dL if they have multiple riskfactors; the recommended LDL:HDL cholesterol ratio in the blood is below5:1, ideally 3.5:1; and the recommended level of free triglycerides inthe blood is less than 200 mg/dL.

Dyslipidemias which the compositions of the present invention are usefulfor preventing or treating hyperlipidemia and low blood levels of highdensity lipoprotein (HDL) cholesterol. In certain embodiments, thehyperlipidemia for prevention or treatment by the compounds of thepresent invention is familial hypercholesterolemia; familial combinedhyperlipidemia; reduced or deficient lipoprotein lipase levels oractivity, including reductions or deficiencies resulting fromlipoprotein lipase mutations; hypertriglyceridemia;hypercholesterolemia; high blood levels of urea bodies (e.g. .beta.-OHbutyric acid); high blood levels of Lp(a) cholesterol; high blood levelsof low density lipoprotein (LDL) cholesterol; high blood levels of verylow density lipoprotein (VLDL) cholesterol and high blood levels ofnon-esterified fatty acids.

The present invention further provides methods for altering lipidmetabolism in a patient, e.g., reducing LDL in the blood of a patient,reducing free triglycerides in the blood of a patient, increasing theratio of HDL to LDL in the blood of a patient, and inhibiting saponifiedand/or non-saponified fatty acid synthesis, said methods comprisingadministering to the patient a compound or a composition comprising afast-acting chromium complex and a slow-acting chromium complex in anamount effective alter lipid metabolism.

Treatment of Dyslipoproteinemias

Also provided are methods for the treatment or prevention of adyslipoproteinemia comprising administering to a patient atherapeutically effective amount of a composition comprising afast-acting chromium complex, a slow-acting chromium complex, and atherapeutically/nutritionally acceptable vehicle.

As used herein, the term “dyslipoproteinemias” refers to disorders thatlead to or are manifested by aberrant levels of circulatinglipoproteins. To the extent that levels of lipoproteins in the blood aretoo high, the compositions described herein are administered to apatient to restore normal levels. Conversely, to the extent that levelsof lipoproteins in the blood are too low, the compositions describedherein are administered to a patient to restore normal levels. Normallevels of lipoproteins are reported in medical treatises known to thoseof skill in the art.

Dyslipoproteinemias which the compositions of the present invention areuseful for preventing or treating include but are not limited to highblood levels of LDL; high blood levels of apolipoprotein B (apo B); highblood levels of Lp(a); high blood levels of apo(a); high blood levels ofVLDL; low blood levels of HDL; reduced or deficient lipoprotein lipaselevels or activity, including reductions or deficiencies resulting fromlipoprotein lipase mutations; hypoalphalipoproteinemia; lipoproteinabnormalities associated with diabetes; lipoprotein abnormalitiesassociated with obesity; lipoprotein abnormalities associated withAlzheimer's Disease; and familial combined hyperlipidemia.

Further provided are methods for reducing apo C-II levels in the bloodof a patient; reducing apo C-III levels in the blood of a patient;elevating the levels of HDL associated proteins, including but notlimited to apo A-I, apo A-II, apo A-IV and apo E in the blood of apatient; elevating the levels of apo E in the blood of a patient, andpromoting clearance of triglycerides from the blood of a patient, saidmethods comprising administering to the patient a fast-acting chromiumcomplex and a slow-acting chromium complex in an amount effective tobring about said reduction, elevation or promotion, respectively.

Treatment of Glucose Metabolism Disorders

Also provided are methods for the treatment or prevention of a glucosemetabolism disorder, comprising providing to a subject with or at riskof developing a glucose metabolism disorder a therapeutically effectiveamount of a fast-acting chromium complex and slow-acting chromiumcomplex and a pharmaceutically acceptable vehicle. As used herein, theterm “glucose metabolism disorders” refers to disorders that lead to orare manifested by aberrant glucose storage and/or utilization. To theextent that indicia of glucose metabolism (i.e., blood insulin, bloodglucose) are too high, the compositions described herein areadministered to a patient to restore normal levels. Conversely, to theextent that indicia of glucose metabolism are too low, the compositionsdescribed herein are administered to a patient to restore normal levels.Normal indicia of glucose metabolism are reported in medical treatisesknown to those of skill in the art.

Glucose metabolism disorders which the compositions of the presentinvention are useful for preventing or treating include but are notlimited to impaired glucose tolerance; insulin resistance; insulinresistance related breast, colon or prostate cancer; diabetes, includingbut not limited to type 2 diabetes, type 1 diabetes, gestationaldiabetes mellitus (GDM), and maturity onset diabetes of the young(MODY); pancreatitis; hypertension; polycystic ovarian disease; HIVlipodystrophy, hormonal imbalance, hypercotisol levers, endothelialdysfunction, Alzheimer's disease, aging and high levels of blood insulinand/or glucose.

Further provided are methods for altering glucose metabolism in apatient, for example to increase insulin sensitivity and/or oxygenconsumption of a patient, said methods comprising administering to themammal a compound or a composition comprising a fast-acting orhydrophilic chromium complex in combination with a slow-acting orlipophilic chromium complex, in an amount effective to alter glucosemetabolism.

Treatment of PPAR-Associated Disorders

Also provided are methods for the treatment or prevention of aPPAR-associated disorder, comprising administering to a patient atherapeutically effective amount of a lipophilic chromium complex and ahydrophilic chromium complex (e.g., chromium histidinate) and apharmaceutically acceptable vehicle. As used herein, “treatment orprevention of PPAR associated disorders” encompasses treatment orprevention of rheumatoid arthritis; multiple sclerosis; psoriasis;inflammatory bowel diseases; breast; colon or prostate cancer; lowlevels of blood HDL; low levels of blood, lymph and/or cerebrospinalfluid apo E; low blood, lymph and/or cerebrospinal fluid levels of apoA-I; high levels of blood VLDL; high levels of blood LDL; high levels ofblood triglyceride; high levels of blood apo B; high levels of blood apoC-III and reduced ratio of post-heparin hepatic lipase to lipoproteinlipase activity. HDL can be elevated in lymph and/or cerebral fluid.

Treatment of Renal Diseases

Further provided are methods for the treatment or prevention of a renaldisease, comprising administering to a patient a therapeuticallyeffective amount of a composition comprising a fast-acting chromiumcomplex and a slow-acting chromium complex and a pharmaceuticallyacceptable vehicle. Renal diseases that can be treated by the compoundsof the present invention include glomerular diseases (including but notlimited to acute and chronic glomerulonephritis, rapidly progressiveglomerulonephritis, nephrotic syndrome, focal proliferativeglomerulonephritis, glomerular lesions associated with systemic disease,such as systemic lupus erythematosus, Goodpasture's syndrome, multiplemyeloma, diabetes, neoplasia, sickle cell disease, and chronicinflammatory diseases), tubular diseases (including but not limited toacute tubular necrosis and acute renal failure, polycystic renaldiseasemedullary sponge kidney, medullary cystic disease, nephrogenicdiabetes, and renal tubular acidosis), tubulointerstitial diseases(including but not limited to pyelonephritis, drug and toxin inducedtubulointerstitial nephritis, hypercalcemic nephropathy, and hypokalemicnephropathy) acute and rapidly progressive renal failure, chronic renalfailure, nephrolithiasis, or tumors (including but not limited to renalcell carcinoma and nephroblastoma). In a most preferred embodiment,renal diseases that are treated by the compounds of the presentinvention are vascular diseases, including but not limited tohypertension, nephrosclerosis, microangiopathic hemolytic anemia,atheroembolic renal disease, diffuse cortical necrosis, and renalinfarcts.

Treatment of Cancer

Provided herein are methods for the treatment or prevention of cancer,comprising administering to a patient a therapeutically effective amountof a composition comprising a fast-acting chromium complex such aschromium histidinate, a slow-acting chromium complex such as chromiumpicolinate, and a pharmaceutically acceptable vehicle. Types of cancerthat can be treated using combination chromium supplementation include,but are not limited to solid tumors, including but not limited tofibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenicsarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma,lymphangioendotheliosarcoma, synovioma mesothelioma, Ewing's tumor,leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer,kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovariancancer, prostate cancer, esophogeal cancer, stomach cancer, oral cancer,nasal cancer, throat cancer, squamous cell carcinoma, basal cellcarcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous glandcarcinoma, papillary carcinoma, papillary adenocarcinomascystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma bile duct carcinoma choriocarcinoma seminoma,embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer,testicular cancer, small cell lung carcinoma, bladder carcinoma, lungcancer, epithelial carcinoma, glioma, glioblastoma multiformeastrocytoma medulloblastoma, craniopharyngioma, ependymoma, pinealoma,hemangioblastoma acoustic neuroma, oligodendroglioma, meningioma, skincancer, melanoma, neuroblastoma, retinoblastoma, Blood-borne cancers,including but not limited to: acute lymphoblastic B-cell leukemia, acutelymphoblastic T-cell leukemia, acute myeloblastic leukemia, “AML,” acutepromyelocytic leukemia “APL,” acute monoblastic leukemia, acuteerythroleukemic leukemia, acute megakaryoblastic leukemia, acutemyelomonocytic leukemia, acute nonlymphocyctic leukemia, acuteundifferentiated leukemia, chronic myelocytic leukemia, “CML,” chroniclymphocytic leukemia, “CLL,” hairy cell leukemia, multiple myeloma Acuteand chronic leukemias, Lymphoblastic myelogenous leukemias, lymphocyticmyelocytic leukemias, Lymphomas: such as Hodgkin's disease,non-Hodgkin's Lymphoma, Multiple myeloma, Waldenstrom'smacroglobulinemia, Heavy chain disease, and Polycythemia vera.

Cancer, including, but not limited to, a tumor, metastasis, or anydisease or disorder characterized by uncontrolled cell growth, can betreated or prevented by combination chromium supplementation asdescribed herein.

Treatment of Other Diseases

Also provided herein are methods for the treatment or prevention ofAlzheimer's Disease, Syndrome X, septicemia, thrombotic disorders,obesity, pancreatitis, hypertension, inflammation, and impotence,comprising administering to a patient a therapeutically effective amountof a composition comprising, consisting essentially of, or consisting ofa fast-acting chromium complex, a slow-acting chromium complex, and anutritionally acceptable vehicle.

As used herein, “treatment or prevention of Alzheimer's Disease”encompasses treatment or prevention of lipoprotein abnormalitiesassociated with Alzheimer's Disease.

As used herein, “treatment or prevention of Syndrome X or MetabolicSyndrome” encompasses treatment or prevention of a symptom thereof,including but not limited to impaired glucose tolerance, hypertensionand dyslipidemia/dyslipoproteinemia.

As used herein, “treatment or prevention of septicemia” encompassestreatment or prevention of septic shock.

As used herein, “treatment or prevention of thrombotic disorders”encompasses treatment or prevention of high blood levels of fibrinogenand promotion of fibrinolysis.

In addition to treating or preventing obesity, the compositionsdescribed herein can be administered to an individual to promote weightreduction of the individual.

As used herein, “treatment or prevention of diabetic nephropathy”encompasses treating or preventing kidney disease that develops as aresult of diabetes mellitus (DM). Diabetes mellitus is a disorder inwhich the body is unable to metabolize carbohydrates (e.g., foodstarches, sugars, cellulose) properly. The disease is characterized byexcessive amounts of sugar in the blood (hyperglycemia) and urine;inadequate production and/or utilization of insulin; and by thirst,hunger, and loss of weight. Thus, the compositions disclosed herein canalso be used to treat or prevent diabetes mellitus.

As used herein, “treatment or prevention of diabetic retinopathy”encompasses treating or preventing complications of diabetes that leadto or cause blindness. Diabetic retinopathy occurs when diabetes damagesthe tiny blood vessels inside the retina, the light-sensitive tissue atthe back of the eye.

As used herein, “treatment or prevention of impotence” includes treatingor preventing erectile dysfunction, which encompasses the repeatedinability to get or keep an erection firm enough for sexual intercourse.The word “impotence” can also be used to describe other problems thatinterfere with sexual intercourse and reproduction, such as lack ofsexual desire and problems with ejaculation or orgasm. The term“treatment or prevention of impotence” includes, but is not limited toimpotence that results as a result of damage to nerves, arteries, smoothmuscles, and fibrous tissues, or as a result of disease, such as, butnot limited to, diabetes, kidney disease, chronic alcoholism, multiplesclerosis, atherosclerosis, vascular disease, and neurologic disease.

As used herein, “treatment or prevention of hypertension” encompassestreating or preventing blood flow through the vessels at a greater thannormal force, which strains the heart; harms the arteries; and increasesthe risk of heart attack, stroke, and kidney problems. The termhypertension includes, but is not limited to, cardiovascular disease,essential hypertension, hyperpiesia, hyperpiesis, malignanthypertension, secondary hypertension, or white-coat hypertension.

As used herein, “treatment or prevention of inflammation” encompassestreating or preventing inflammation diseases including, but not limitedto, chronic inflammatory disorders of the joints including arthritis,e.g., rheumatoid arthritis and osteoarthritis; respiratory distresssyndrome, inflammatory bowel diseases such as ileitis, ulcerativecolitis and Crohn's disease; and inflammatory lung disorders such asasthma and chronic obstructive airway disease, inflammatory disorders ofthe eye such as corneal dystrophy, trachoma, onchocerciasis, uveitis,sympathetic ophthalmitis, and endophthalmitis; inflammatory disorders ofthe gum, e.g., periodontitis and gingivitis; tuberculosis; leprosy;inflammatory diseases of the kidney including glomerulonephritis andnephrosis; inflammatory disorders of the skin including acne,sclerodermatitis, psoriasis, eczema, photoaging and wrinkles;inflammatory diseases of the central nervous system, includingAIDS-related neurodegeneration, stroke, neurotrauma, Alzheimer'sdisease, encephalomyelitis and viral or autoimmune encephalitis;autoimmune diseases including immune-complex vasculitis, systemic lupusand erythematodes; systemic lupus erythematosus (SLE); and inflammatorydiseases of the heart such as cardiomyopathy.

Combination Therapy

In certain embodiments, the compounds and compositions disclosed hereincan be used in combination therapy with at least one other therapeuticagent. The combination chromium supplementation described herein (e.g.,a fast-acting chromium complex and a slow-acting chromium complex) andthe therapeutic agent can act additively or, more preferably,synergistically. In a preferred embodiment, combination chromiumsupplementation described herein is administered concurrently with theadministration of another therapeutic agent, which can be part of thesame combination chromium supplementation described herein or adifferent composition. In another embodiment, both a hydrophilicchromium complex and a lipophilic chromium complex are administeredprior or subsequent to administration of another therapeutic agent. Asmany of the disorders for which the compositions described herein areuseful in treating are chronic disorders, in one embodiment combinationtherapy involves alternating between administering a combinationchromium supplementation and a composition comprising anothertherapeutic agent, e.g., to minimize the toxicity associated with aparticular drug. The duration of administration of each drug ortherapeutic agent can be, e.g., one month, three months, six months, ora year. In certain embodiments, when a composition described herein isadministered concurrently with another therapeutic agent thatpotentially produces adverse side effects including but not limited totoxicity, the therapeutic agent can advantageously be administered at adose that falls below the threshold at which the adverse side iselicited. The standard dosage for the compounds discussed below areknown to those skilled in the art.

The combination chromium supplementation can be administered togetherwith a statin. Statins for use in combination with the compounds andcompositions described herein include but are not limited toatorvastatin, pravastatin, fluvastatin, lovastatin, simvastatin, andcerivastatin.

The combination chromium supplementation can also be administeredtogether with a PPAR agonist, for example a thiazolidinedione or afibrate.

Thiazolidinediones for use in combination with the compounds andcompositions described herein include but are not limited to 5 ((4 (2(methyl 2 pyridinylamino)ethoxy)phenyl)methyl) 2,4 thiazolidinedione,troglitazone, pioglitazone, ciglitazone, WAY 120,744, englitazone, AD5075, darglitazone, and rosiglitazone. Fibrates for use in combinationwith the compounds and compositions described herein include but are notlimited to gemfibrozil, fenofibrate, clofibrate, or ciprofibrate. Asmentioned previously, a therapeutically effective amount of a fibrate orthiazolidinedione often has toxic side effects. Accordingly, in apreferred embodiment of the present invention, when a combinationchromium supplementation is administered in combination with a PPARagonist, the dosage of the PPAR agonist is below that which isaccompanied by toxic side effects.

The combination chromium supplementation can also be administeredtogether with a bile acid binding resin. Bile acid binding resins foruse in combination with the compounds and compositions described hereininclude but are not limited to cholestyramine and colestipolhydrochloride. The present compositions can also be administeredtogether with niacin or nicotinic acid. The present compositions canalso be administered together with a RXR agonist. RXR agonists for usein combination with the compositions disclosed herein include but arenot limited to LG 100268, LGD 1069, 9-cis retinoic acid, 2 (1 (3,5,5,8,8pentamethyl 5,6,7,8 tetrahydro 2 naphthyl) cyclopropyl) pyridine 5carboxylic acid, or 4 ((3,5,5,8,8 pentamethyl 5,6,7,8 tetrahydro 2naphthyl)2 carbonyl) benzoic acid. The combination chromiumsupplementation can also be administered together with an anti-obesitydrug. Anti-obesity drugs for use in combination with the combinationchromium supplementation disclosed herein include but are not limited toβ.-adrenergic receptor agonists, preferably β.-3 receptor agonists,fenfluramine, dexfenfluramine, sibutramine, bupropion, fluoxetine, andphentermine. The combination chromium supplementation can also beadministered together with a hormone. Hormones for use in combinationwith the compositions disclosed herein include but are not limited tothyroid hormone, estrogen and insulin. Preferred insulins include butare not limited to injectable insulin, transdermal insulin, inhaledinsulin, or any combination thereof. As an alternative to insulin, aninsulin derivative, secretagogue, sensitizer or mimetic can be used.Insulin secretagogues for use in combination with the combinationchromium supplementation disclosed herein include but are not limited toforskolin, dibutryl cAMP or isobutylmethylxanthine (IBMX).

The combination chromium supplementation can also be administeredtogether with a phosphodiesterase type 5 (“PDE5”) inhibitor to treat orprevent disorders, such as but not limited to, impotence. In aparticular, embodiment the combination is a synergistic combination of acombination chromium supplementation and a PDE5 inhibitor.

The present compositions can also be administered together with atyrophostine or an analog thereof. Tyrophostines for use in combinationwith the combination chromium supplementation include but are notlimited to tryophostine 51.

The present compositions can also be administered together withsulfonylurea-based drugs. Sulfonylurea-based drugs for use incombination chromium supplementation disclosed herein include, but arenot limited to, glisoxepid, glyburide, acetohexamide, chlorpropamide,glibornuride, tolbutamide, tolazamide, glipizide, gliclazide,gliquidone, glyhexamide, phenbutamide, and tolcyclamide. The combinationchromium supplementation can also be administered together with abiguanide. Biguanides for use in combination with the compositionsdisclosed herein include but are not limited to metformin, phenforminand buformin.

The combination chromium supplementation can also be administeredtogether with an α-glucosidase inhibitor. α-glucosidase inhibitors foruse in combination with the combination chromium supplementation includebut are not limited to acarbose and miglitol.

The combination chromium supplementation can also be administeredtogether with an apo A-I agonist. In one embodiment, the apo A-I agonistis the Milano form of apo A-I (apo A-IM). In a preferred mode of theembodiment, the apo A-IM for administration in conjunction with thecombination chromium supplementation is produced by the method of U.S.Pat. No. 5,721,114 to Abrahamsen. In a more preferred embodiment, theapo A-I agonist is a peptide agonist. In a preferred mode of theembodiment, the apo A-I peptide agonist for administration inconjunction with the combination chromium supplementaiton is a peptideof U.S. Pat. No. 6,004,925 or 6,037,323 to Dasseux.

The combination chromium supplementation can also be administeredtogether with apolipoprotein E (apo E).

In yet other embodiments, the combination chromium supplementation canbe administered together with an HDL-raising drug; an HDL enhancer; or aregulator of the apolipoprotein A-I, apolipoprotein A-IV and/orapolipoprotein genes.

In one embodiment, the other therapeutic agent can be an antiemeticagent. Suitable antiemetic agents include, but are not limited to,metoclopromide, domperidone, prochlorperazine, promethazine,chlorpromazine, trimethobenzamide, ondansetron, granisetron,hydroxyzine, acethylleucine monoethanolamine, alizapride, azasetron,benzquinamide, bietanautine, bromopride, buclizine, clebopride,cyclizine, dimenhydrinate, diphenidol, dolasetron, meclizine,methallatal, metopimazine, nabilone, oxyperndyl, pipamazine,scopolamine, sulpiride, tetrahydrocannabinols, thiethylperazine,thioproperazine and tropisetron.

In another embodiment, the other therapeutic agent can be anhematopoietic colony stimulating factor. Suitable hematopoietic colonystimulating factors include, but are not limited to, filgrastim,sargramostim, molgramostim and erythropoietin a.

In still another embodiment, the other therapeutic agent can be anopioid or non-opioid analgesic agent. Suitable opioid analgesic agentsinclude, but are not limited to, morphine, heroin, hydromorphone,hydrocodone, oxymorphone, oxycodone, metopon, apomorphine, normorphine,etorphine, buprenorphine, meperidine, lopermide, anileridine,ethoheptazine, piminidine, betaprodine, diphenoxylate, fentanil,sufentanil, alfentanil, remifentanil, levorphanol, dextromethorphan,phenazocine, pentazocine, cyclazocine, methadone, isomethadone andpropoxyphene. Suitable non-opioid analgesic agents include, but are notlimited to, aspirin, celecoxib, rofecoxib, diclofinac, diflusinal,etodolac, fenoprofen, flurbiprofen, ibuprofen, ketoprofen, indomethacin,ketorolac, meclofenamate, mefanamic acid, nabumetone, naproxen,piroxicam and sulindac.

Combination Therapy of Cardiovascular Diseases

The combination chromium supplementation (e.g., slow-acting chromiumcomplex and fast-acting chromium complex) can be administered togetherwith a known cardiovascular drug. Cardiovascular drugs for use incombination with the combination chromium supplementation to prevent ortreat cardiovascular diseases include but are not limited to peripheralantiadrenergic drugs, centrally acting antihypertensive drugs (e.g.,methyldopa, methyldopa HCl), antihypertensive direct vasodilators (e.g.,diazoxide, hydralazine HCl), drugs affecting renin-angiotensin system,peripheral vasodilators, phentolamine, antianginal drugs, cardiacglycosides, inodilators (e.g., aminone, milrinone, enoximone,fenoximone, imazodan, sulmazole), antidysrhythmic drugs, calcium entryblockers, ranitine, bosentan, and rezulin.

Surgical Uses

Cardiovascular diseases such as atherosclerosis often require surgicalprocedures such as angioplasty. Angioplasty is often accompanied by theplacement of a reinforcing a metallic tube shaped structure known as a“stent” into a damaged coronary artery. For more serious conditions,open heart surgery such as coronary bypass surgery may be required.These surgical procedures entail using invasive surgical devices and/orimplants, and are associated with a high risk of restenosis andthrombosis. Accordingly, the combination chromium supplementationdescribed herein (i.e., a lipophilic chromium complex and a hydrophilicchromium complex) can be used as coatings on surgical devices (e.g.,catheters) and implants (e.g., stents) to reduce the risk of restenosisand thrombosis associated with invasive procedures used in the treatmentof cardiovascular diseases.

Veterinary and Livestock Uses

The combination chromium supplementation (i.e., a lipophilic chromiumcomplex and a hydrophilic chromium complex) can be administered to ananimal or non-human animal for a veterinary use for treating orpreventing a disease or disorder disclosed herein.

In a specific embodiment, the non-human animal is a household pet. Inanother specific embodiment, the non-human animal is a livestock animal.In a preferred embodiment, the non-human animal is a mammal, mostpreferably a cow, horse, sheep, pig, cat, dog, mouse, rat, rabbit, orguinea pig. In another preferred embodiment, the non-human animal is afowl species, most preferably a chicken, turkey, duck, goose, or quail.

In addition to veterinary uses, the combination chromium supplementationdisclosed herein can be used to reduce the fat content of livestock toproduce leaner meats. Alternatively, the combination chromiumsupplementation disclosed herein can be used to reduce the cholesterolcontent of eggs by administering the compounds to a chicken, quail, orduck hen. For non-human animal uses, the compositions disclosed hereincan be administered via the animals' feed or orally as a drenchcomposition.

Therapeutic/Prophylactic Administration and Compositions

Due to the activity of the combination chromium supplementationdescribed herein, they are useful in veterinary and human medicine. Asdescribed above, the combination chromium supplementation describedherein are useful for the treatment or prevention of cardiometabolicsyndrome, aging, Alzheimer's Disease, cancer, cardiovascular disease,diabetic nephropathy, diabetic retinopathy, a disorder of glucosemetabolism, dyslipidemia, dyslipoproteinemia, hypertension, impotence,inflammation, insulin resistance, lipid elimination in bile, modulatingC reactive protein, obesity, oxysterol elimination in bile,pancreatitis, Parkinson's disease, a peroxisome proliferator activatedreceptor-associated disorder, phospholipid elimination in bile, renaldisease, septicemia, metabolic syndrome disorders (e.g., Syndrome X), athrombotic disorder, enhancing bile production,-enhancing reverse lipidtransport, inflammatory processes and diseases like gastrointestinaldisease, irritable bowel syndrome (IBS), inflammatory bowel disease(e.g., Crohn's Disease, ulcerative colitis), arthritis (e.g., rheumatoidarthritis, osteoarthritis), autoimmune disease (e.g., systemic lupuserythematosus), scleroderma, ankylosing spondylitis, gout andpseudogout, muscle pain: polymyositis/polymyalgia rheumatica/fibrositis;infection and arthritis, juvenile rheumatoid arthritis, tendonitis,bursitis and other soft tissue rheumatism.

Provided herein are methods of treatment and prophylaxis of theconditions enumerated above by providing to a subject of atherapeutically effective amount of a fast-acting chromium complex and aslow-acting chromium complex as disclosed herein. In the variousembodiments disclosed herein, the subject can be a mammal such as ananimal, including, but not limited, to an animal such a cow, horse,sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit, guineapig, etc., and most preferably a human.

The combination chromium supplementation disclosed herein is useful formethods for treating diabetes and its related pathologies,cardiovascular and related diseases, such as, for example, diabetesretinopathy, diabetes nephropathy, diabetes neuropathy, diabetes footproblems, diabetes infections and inflammations, diabetes withcardiovascular complications such as hypertrophy, hypertension,congestive heart failure, myocardial ischemia, ischemia reperfusioninjuries in an organ, arrhythmia, and myocardial infarction. Oneembodiment is directed to a method of treating cardiovascular disease ina mammal by concurrently administering to the mammal a therapeuticallyeffective amount of a combination chromium supplementation describedherein (e.g. a lipophilic chromium complex and a hydrophilic chromiumcomplex) and a therapeutic cardiovascular compound such as chromiumhistidine or chromium complex. Therapeutic cardiovascular compoundssuitable for use in methods described herein include an angiotensinconverting enzyme inhibitor, an angiotensin II receptor antagonist, acalcium channel blocker, an anti-thrombotic agent, a β-adrenergicreceptor antagonist, a vasodilator, a diuretic, an α-adrenergic receptorantagonist, an antioxidant, and a mixture thereof.

The combination chromium supplementation disclosed herein is useful forthe methods for treating obesity and related pathologies, obesityrelated to complications such as diabetes, diabetes risk factors, leptinresistance, abdominal fat distribution, cardiovascular disease and itsrelated pathologies, cardiovascular and related diseases, such as, forexample, hypertrophy, hypertension, congestive heart failure, myocardialischemia, ischemia reperfusion injuries in an organ, arrhythmia, andmyocardial infarction. One embodiment is directed to a method oftreating obesity and its associated complications such as diabetes,cardiovascular disease and insulin resistance in a mammal byconcurrently administering to the mammal a therapeutically effectiveamount of a combination chromium suppplementation (e.g. a lipophilicchromium complex and a hydrophilic chromium complex) and a therapeuticcardiovascular compound. Therapeutic cardiovascular compounds caninclude but are not limited to an angiotensin converting enzymeinhibitor, an angiotensin II receptor antagonist, a calcium channelblocker, an anti-thrombotic agent, a β-adrenergic receptor antagonist, avasodilator, a diuretic, an α-adrenergic receptor antagonist, anantioxidant, antihyperglycemic drugs, insulin, antiobesity drugs,antidepressants etc. and a mixture thereof.

The present invention is further disclosed in the following Examples,which are provided for illustrative purposes and are not in any wayintended to limit the scope of the invention as claimed.

Example 1 Combination Chromium Picolinate/Chromium Histidinate ImprovesGlucose Uptake In Vitro

The following example describes experiments that compared the effects ofchromium supplementation in the form of chromium picolinate versus acombination of chromium picolinate:chormium histidinate (1:1 elementalchromium) on glucose uptake in skeletal muscle cells. The amounts ofchromium picolinate and the combination were adjusted so as to provideequimolar amounts of elemental chromium. For the combination, 50% of theelemental chromium was provided in the form of chromium picolinate, and50% was provided in the form of chromium histidinate. To assess glucoseuptake, [³H]C-deoxyglucose was added to cultures of skeletal muscle, inmedia with or without insulin and with or without the indicated chromiumsupplementation. Uptake of the labeled glucose by the cells wascalculated.

FIG. 1 shows the percent increase in glucose uptake in culturesreceiving chromium supplementation over cultures without chromiumsupplementation, in the presence and absence of insulin. Both chromiumpicolinate and the combination chromium picolinate/chromium histidinatestimulated glucose uptake in insulin-stimulated cells. The combinationenhanced glucose uptake in both basal and insulin-stimulated cultures toa greater extent than chromium picolinate, even though the cultures weretreated with equivalent amounts of elemental chromium.

Example 2 Combination Chromium Picolinate/Chromium HistidinateSynergistically Improves Glucose Levels, Cholesterol Levels,Triglyceride Levels and Free Fatty Acid Levels In Vivo

The following example describes a series of experiments that comparedthe effects of chromium supplementation in the form of chromiumpicolinate, chromium histidinate, and a combination of chromiumpicolinate/chromium histidinate on various metabolic functions in ratsfed normal and high fat diets.

Wistar rats were reared at the temperature of (22±2° C.), humidity(55±5%) and a 12/12 h light/dark cycle. Pellet food and water wereprovided ad libitum.

Rats were randomly assigned to one of eight treatment groups.

Group 1: Control rats were fed standard diet (12% of calories as fat)for 12 weeks.

Group 2: Rats were fed standard diet with chromium picolinate (14.3 μg,providing 1.43 μg chromium) included in the water for 12 weeks.

Group 3: Rats were fed standard diet with chromium histidinate (19.7 μg,providing 1.43 μg chromium) included in the water for 12 weeks.

Group 4: Rats were fed standard diet with chromium picolinate (7.15 μg)and chromium histidinate (9.85 μg), providing 1.43 μg total chromium inthe water for 12 weeks.

Group 5: Rats were fed high fat diet (40% of calories as fat) for 12weeks.

Group 6: Rats were fed high-fat diet (40% of calories as fat) and withchromium picolinate (14.3 μg, providing 1.43 μg chromium) included inthe water for 12 weeks.

Group 7: Rats were fed high-fat diet (40% of calories as fat) and withchromium histidinate (19.7 μg, providing 1.43 μg chromium) included inthe water for 12 weeks.

Group 8: Rats were fed high-fat diet (40% of calories as fat) and withchromium picolinate (7.15 μg) and chromium histidinate (9.85 m),providing 1.43 μg total chromium in the water for 12 weeks.

Bodyweight, serum glucose, composite insulin sensitivity index (CISI),serum cholesterol, serum triglycerides and serum free fatty acid levelswere measured. The baseline levels for the control rats (no chromiumsupplementation) that were fed normal or high fat diets are shown inTable 1, below.

TABLE 1 Normal Rats High Fat Diet Rats Body weight (g) 277 317 Serumglucose (mg/dL) 100 130 CISI (mg/dL) 2.7 1.6 Serum cholesterol (mg/dL)58 112 Serum triglycerides (mg/dL) 46 107 Free fatty acids (mmol/L) 1.43.1

Serum glucose levels, serum cholesterol levels, serum triglyceridelevels, and free fatty acid levels were measured in all groups aftertwelve weeks. The data are presented in FIGS. 2A-5B.

Chromium histidinate decreased serum glucose levels in both normal andhigh-fat fed rats to a significantly greater degree than chromiumpicolinate. Surprisingly, the treatment group that received chromiumpicolinate/chromium histidinate (in the same total amount of elementalchromium as provided by chromium picolinate or chromium histidinatealone) exhibited a significantly greater reduction in serum glucoselevels than the treatment groups that received chromium supplementationin the form of chromium as chromium picolinate or chromium histidinatealone. FIGS. 2A and 2B. The synergistic effect of the combination couldnot have been predicted.

Chromium picolinate, chromium histidinate and the combination ofchromium picolinate/chromium histidinate resulted in a significantdecrease in total serum cholesterol levels in rats fed a normal diet.FIG. 3A. Surprisingly, the treatment group that received chromiumpicolinate/chromium histidinate (in the same total amount of elementalchromium as provided by chromium picolinate or chromium histidinatealone) had significantly lower serum cholesterol levels than thetreatment groups that received chromium supplementation in the form ofchromium as chromium picolinate or chromium histidinate alone. FIG. 3B.The synergistic effect of the combination could not have been predicted.

Chromium picolinate, chromium histidinate and the combination ofchromium picolinate/chromium histidinate resulted in a significantdecrease in serum triglyceride levels in rats fed a normal or high fatdiet. FIGS. 4A and 4B. Surprisingly, the treatment group that receivedchromium picolinate/chromium histidinate (in the same total amount ofelemental chromium as provided by chromium picolinate or chromiumhistidinate alone) had significantly lower serum triglyceride levelsthan the treatment groups that received chromium supplementation in theform of chromium as chromium picolinate or chromium histidinate alone.FIG. 4B. The synergistic effect of the combination could not have beenpredicted.

Chromium picolinate, chromium histidinate and the combination ofchromium picolinate/chromium histidinate resulted in a significantdecrease in free fatty acid levels in rats fed a normal or high fatdiet. FIGS. 5A and 5B. Surprisingly, the treatment group that receivedchromium picolinate/chromium histidinate (in the same total amount ofelemental chromium as provided by chromium picolinate or chromiumhistidinate alone) had significantly lower free fatty acid levels thanthe treatment groups that received chromium supplementation in the formof chromium as chromium picolinate or chromium histidinate alone. FIG.5B. The synergistic effect of the combination could not have beenpredicted.

Example 3 Combination Chromium Supplementation to Achieve SustainedRelease of Chromium

Combination supplementation with a fast-acting chromium complex and aslow-acting chromium complex results in a sustained release of chromiumover a period of time. More specifically, the combination chromiumsupplementation is useful in achieving prolonged, sustained chromiumabsorption to an individual in need thereof. Chromium supplementationhas many therapeutic indications. For example, chromium has beenassociated with the improvement of glucose metabolism, mitigation ofinsulin resistance, improvement of symptoms associated withhyperglycemia, improvement of lipid profiles, promotion of lean bodymass, and the reduction of obesity.

One of the surprising aspects of the present invention is the discoverythat the combination chromium supplementation with a hydrophilicchromium complex and lipophilic chromium complex results in a greaterthan additive effect in ameliorating some of the diseases, conditions,and associated symptoms disclosed above. Without being bound to aparticular theory, it is believed that the synergistic effects observedwith combination chromium therapy can be attributed to the achievementof rapid chromium absorption via the hydrophilic chromium complexsupplementation and sustained, long-term absorption via the lipophilicchromium complex supplementation. By combining a fast-acting chromiumcomplex with a slow-acting chromium complex, prolonged, sustainedabsorption of chromium by an individual in need thereof is accomplished.

An individual presenting with poor glucose metabolism is identified(e.g., either self-identified or identified by other means). Theindividual is administered a nutritional supplement comprising aneffective dose of chromium histidinate and an effective dose of chromiumpicolinate. Glucose metabolism is monitored and an improvement inglucose metabolism is observed.

An individual with high LDL cholesterol and low HDL cholesterol isidentified (e.g., either self-identified or identified by other means).The individual is administered an effective dose of chromium acetate andan effective dose of chromium picolinate. An improvement of theindividual's lipid profile is observed. LDL cholesterol is lowered andHDL cholesterol is raised.

An individual with insulin resistance is identified (e.g., eitherself-identified or identified by other means). The individual isadministered a dietary beverage comprising an effective dose of chromiumhistidinate and an effective dose of chromium picolinate. Theindividual's response to insulin is monitored. The supplementation withchromium histidinate and chromium picolinate results in an improvementin insulin sensitivity.

The methods, compositions, and devices described herein are presentlyrepresentative of preferred embodiments and are exemplary and are notintended as limitations on the scope of the invention. Changes thereinand other uses will occur to those skilled in the art which areencompassed within the spirit of the invention and are defined by thescope of the disclosure. Accordingly, it will be apparent to one skilledin the art that varying substitutions and modifications can be made tothe invention disclosed herein without departing from the scope andspirit of the invention.

As used in the claims below and throughout this disclosure, by thephrase “consisting essentially of” is meant including any elementslisted after the phrase, and limited to other elements that do notinterfere with or contribute to the activity or action specified in thedisclosure for the listed elements. Thus, the phrase “consistingessentially of” indicates that the listed elements are required ormandatory, but that other elements are optional and may or may not bepresent depending upon whether or not they affect the activity or actionof the listed elements.

Numerous literature and patent references have been cited in the presentpatent application. Each and every reference that is cited in thispatent application is incorporated by reference herein in its entirety.

What is claimed is:
 1. A composition comprising: a chromium complexcomprising: an amount of a fast-acting chromium complex; and an amountof a slow-acting chromium complex; wherein the amount of a fast actingchromium complex and the amount of the slow-reacting chromium complexare formulated to have a molar ratio of chromium between 1:10 and 10:1.2. The composition of claim 1, wherein the chromium complex is ahydrophobic chromium complex.
 3. The composition of claim 1, wherein thefast-acting chromium complex is hydrophilic and the slow-acting chromiumcomplex is lipophilic.
 4. The composition of claim 1, wherein theslow-acting chromium complex is lipophilic.
 5. The composition of claim1, wherein the fast-acting chromium complex is hydrophilic.
 6. Thecomposition of claim 1, wherein the amount of the fast-acting chromiumcomplex and the amount of the slow-acting chromium complex areformulated to have a molar ratio of chromium between 1:8 and 8:1.
 7. Thecomposition of claim 1, wherein the amount of the fast-acting chromiumcomplex and the amount of the slow-acting chromium complex areformulated to have a molar ratio of chromium between 1:4 and 4:1.
 8. Thecomposition of claim 1, wherein the amount of the fast-acting chromiumcomplex and the amount of the slow-acting chromium complex areformulated to have a molar ratio of chromium between 1:2 and 2:1.
 9. Thecomposition of claim 1, wherein the amount of the fast-acting chromiumcomplex and the amount of the slow-acting chromium complex areformulated to have a molar ratio of chromium of about 1:1.
 10. Thecomposition of claim 1, wherein the amount of a fast-acting chromiumcomplex comprises a chromium histidinate complex.
 11. The composition ofclaim 1, wherein the amount of a fast-acting chromium complex consistsessentially of a chromium histidinate complex.
 12. The composition ofclaim 1, wherein the amount of a fast-acting chromium complex comprises,chromium acetate, chromium chloride, chromium histidinate, and chromiumnicotinate, or any combination thereof.
 13. The composition of claim 1,wherein the first amount of a fast-acting chromium complex compriseschromium histidinate, chromium trihistidinate, chromium polyhistidinate,or any combination thereof.
 14. The composition of claim 1, furthercomprising metformin.
 15. The composition of claim 1, further comprisinga pharmaceutically acceptable vehicle.
 16. The composition of claim 1,wherein the slow-acting chromium complex comprises chromium picolinate,chromium tripicolinate, or a combination thereof.
 17. The composition ofclaim 1, wherein the slow-acting chromium complex comprises chromiumpicolinate.
 18. The composition of claim 1, wherein the slow-actingchromium complex comprises chromium tripicolinate.
 19. The compositionof claim 1, wherein the slow-acting chromium complex provides a glucoselowering effect.
 20. The composition of claim 1, wherein the slow-actingchromium complex provides a glucose stabilization effect for at least 10hours.